In recent years, with the development of metabolic reprogramming research, people have changed their understanding of the biological effects of immune cells. Under the stimulation of inflammatory response, immune cells re-regulate their metabolism and bioenergetics, provide energy and substrates for cell survival, and initiate immune effect functions. Nod-like receptor protein 3 (NLRP3) inflammasome, as an important component of the innate immune system, has been shown to sense metabolites such as uric acid and cholesterol crystals, and can be inhibited by metabolites such as ketones. It is also regulated by mitochondrial reactive oxygen species and glycolytic components (such as hexokinase). Recent studies have shown that a variety of metabolic pathways converge as effective regulators of NLRP3 inflammasome. The paper reviews the metabolic regulatory pathways and specificity of NLRP3 inflammasome activation, and its role in renal diseases.

Neoadjuvant therapy has become the first choice for locally advanced esophageal carcinoma. Patients with post-neoadjuvant positive lymph node staging (ypN+) have poor prognosis, and there is no effective adjuvant therapy. Programmed death protein-1 (PD-1) antibody can obtain better clinical efficacy in the treatment of advanced esophageal cancer. The authors designed a multicenter, prospective, randomized controlled clinical trial of Toripalimab (PD-1 antibody) adjuvant therapy on esophageal squamous cell carcinoma patients with ypN+ after the treatment of neoadjuvant chemotherapy combined with surgical resection, in order to provide clinical practices for the adjuvant treatment of ypN+ patients.

Objective To investigate the expression and mechanism of microRNA-148b (miRNA-148b) in high glucose-induced renal tubular injury.Method HK-2 cells cultured in vitro were divided into normal glucose group,mannitol hypertonic control group and high glucose group.After 48 hours of culture,the expression of miRNA-148b was detected by real-time quantitative PCR.2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) was used for detecting production of ROS and observed under fluorescence microscope for analysis;The expression of AMPKot1,Bcl-2,NOX2,NOX4,activated caspase3 (cleaved-caspase3) were detected by Western blotting.Results Compared with the normal glucose group,the expression of miRNA-148b was up-regulated in HK-2 cells in high glucose group and hypertonic group (P ＜ 0.01),and the production of ROS increased (P ＜ 0.01).The expression of NOX2 and NOX4 was increased,AMPKα1 and Bcl-2 decreased,and cleaved caspase-3 was increased (all P ＜ 0.01).Conclusions HG up-regulated miRNA-148b expression and down-regulated its target gene AMPKα1 which promotes the expression of NOX2 and NOX4 in HK-2 cells.MiRNA-148b promotes apoptosis of HK-2 cells via increasing production of ROS and enhancing cleaved-caspase3 for Bcl-2 insufficiency.The tubular toxicity of high glucose is partly due to osmotic pressure.MiRNA-148b may be involved in the pathological injury of diabetic nephropathy and is expected to become a new therapeutic target for diabetic nephropathy.

Objective To investigate whether advanced glycation end products (AGEs) can induce the expression of Ros,JC-1 and its apoptosis-related proteins in glomerular mesangial cells under high glucose environment,induce apoptosis and injury of glomerular mesangial cells.Methods Rat glomerular mesangial cell line HBZY-1 was cultured in vitro.The cells were cultured with different concentrations of AGEs for 0,12,24 and 48 hours respectively.MTT assay was used to observe the cell proliferation ability.After the optimal time and concentration of AGEs were selected,the caspase enzyme inhibitor Z-VAD-fmk and reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine (NAC) were cultured and the apoptosis rate was detected by cell death detection apoptosis ELISA plus and Annexin V-FITC/PI kit.JC-1 staining was used to detect the changes of mitochondrial membrane potential (MMP).Cell ROX deep red flow cytometry was used to detect the total ROS level.The expression of anti-apoptotic protein Bcl-2,pro-apoptotic protein BAX,caspase-9,caspase-3 and poly ADP-ribose polymerase (PARP)-activated fragments was detected by Western blotting.Results AGEs could decrease the activity of glomerular mesangial cells in a time and concentration-dependent manner,and induce cell death.The percentage of apoptotic cells in glomerular mesangial cells was significantly increased after treatment with 250 mg/L AGEs for 24 h (P ＜ 0.01),and Z-VAD-fmk could significantly alleviate AGEs-induced glomerular mesangial cell apoptosis (P ＜ 0.01).Compared with the control group,AGEs increased the level of intracellular reactive oxygen species and decreased MMP in a time-dependent manner,and the two time points that AGEs significantly caused the change were 1 h and 2 h (all P ＜ 0.01).AGEs also reduced the expression of antiapoptotic protein Bcl-2 and increased the expression of pro-apoptotic protein Bax,cleaved caspase-9,cleaved caspase-3 and cleaved PARP (all P ＜ 0.01).Compared with AGEs group,NAC could significantly stabilize MMP (P ＜ 0.01),increase Bcl-2 expression (P ＜ 0.01),and decrease the expression of BAX,cleaved caspase-9,cleaved caspase-3 and cleaved PARP (all P ＜ 0.01).Conclusion AGEs induce mitochondrial pathway apoptosis in glomerular mesangial cells by increasing intracellular ROS level and destroying MMP.

Objective To detect the serum microRNA-148b-3p level in patients with diabetes mellitus and diabetic nephropathy,and to analyze its correlation with clinical and pathological indexes.Methods The research crowd was divided into three groups (1) diabetic nephropathy group:biopsy with diabetic nephropathy (n=25,14 males,11 females);(2) type 2 diabetes mellitus group:type 2 diabetes mellitus patients with normal urinary microalbumin/urinary creatinine value (n=10,4 males,6 females);(3) normal control group:healthy subjects (n=9,3 males and 6 females).Clinical indicators included gender,age,24-hour urine protein,systolic blood pressure (SBP),diastolic blood pressure (DBP),serum creatinine (Scr),urea (Urea),cystatin-C (Cys-C),blood albumin (ALB),urine microalbumin (UMA),triacylglycerol (TG),total cholesterol (TC),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),serum uric acid (UA),fasting blood glucose (FBG),glycated hemoglobin (HbA1c),urine microalbuminuria / urinary creatinine (UACR),and estimated glomerular filtration rate (eGFR) calculated by CKD-EPI formula.Real-time quantitative PCR was applied to verify the expression of microRNA-148b-3p in serum samples of the research crowds.The relationships between microRNA-148b-3p level and clinical features was also analyzed.Results The levels of serum microRNA-148b-3p in diabetic nephropathy group and in type 2 diabetes mellitus group were 1.82 times and 1.73 times of that in normal control group (P ＜ 0.05,respectively).The level of serum microRNA-148b-3p was significantly correlated with HDL-C (r=-0.374,P=0.013),UMA (r=0.426,P=0.004),FBG (r=0.330,P=0.046) and TG (r=0.423,P=0.005).Multiple linear regression analysis showed that UMA level was independently associated with serum microRNA-148b-3p level (β=0.338,P=0.044).The area under the receiver operating characteristic curve (ROC) of serum microRNA-148b-3p in diagnosing type 2 diabetes mellitus and diabetic nephropathy was 0.835 and 0.665,respectively.Conclusions The level of serum microRNA-148b-3p of patients with type 2 diabetes mellitus or diabetic nephropathy significantly increases.The level of UMA is independently associated with serum microRNA-148b-3p level.Serum microRNA-148b-3p is expected to be a potential biomarker for the diagnosis of diabetic nephropathy.

Objective: To investigate the effects of abated microRNA-21 (miRNA-21) on phosphatase and tensin homologue on chromosome ten protein (PTEN) and PI3K/Akt/mTOR pathway, as well as their further influence on the autophagy in high glucose (HG, 25.0 mmol/L) induced rat glomerular mesangial cells. Methods: MiRNA-21 inhibitor and negative control were transfected by liposome 2000 into rat glomerular mesangial cells (HBZY-1). The cells were divided into normal glucose (5.5 mmol/L) group, normal glucose+negative control group, normal glucose+miRNA-21 inhibitor group, HG group, HG+negative control group and HG+miRNA-21 inhibitor group. Cell proliferation and hypertrophy were assayed by MTT and the ratio of total protein to cell number respectively. The miRNA-21 expression was detected using real time PCR. The expressions of PTEN/Akt/mTOR signaling signatures, autophagy-associated protein (p62 and LC3 Ⅱ) and collagen Ⅰ was detected by Western blotting and real time PCR. Autophagosomes were observed using electron microscopy. Results: Compared with those in normal glucose group, in HG group cells had hypertrophy and proliferation, up-regulated miRNA-21 expression, and down-regulated PTEN protein and mRNA expressions (all P<0.01). Also there were and up-regulated p-Akt, p-mTOR, p62 and collagen Ⅰ expression, and lower LC3 Ⅱ expression and autophagosomes (all P<0.01). Further, compared with those in HG group, cells hypertrophy and proliferation in HG+miRNA-21 inhibitor group were reduced, expressions of p-Akt, p-mTOR, p62 and collagen Ⅰ were down-regulated, while expressions of PTEN and LC3 Ⅱ and autophagosomes were up-regulated (all P<0.01). Conclusions MiRNA-21 inhibitor up-regulates PTEN expression, which inhibits the activation of Akt/mTOR signaling pathway, ameliorates cell hypertrophy, proliferation and enhances autophagy to reduce extracellular matrix accumulation.

Objective To investigate whether the effect of transient high glucose on inflammatory factors expression could be continuous in rat glomerular mesangial cell,and its relation with histone methylation modification.Methods Rat glomerular mesangial cells (HBZY-l) were divided into three groups:the high glucose group (25.0 mmol/L glucose),the hypertonic group (MA,5.5 mmol/L glucose+ 19.5 mmol/L mannitol) and the normal-glucose control group (5.5 mmol/L glucose),which were cultured for 24 h respectively.All 3 groups were then changed with normal-glucose medium to culture for 24 h,48 h and 72 h.Their protein,mRNA and supernatant were harvested.The protein expressions of mono-methylation of H3 lysine 4 (H3K4mel) was measured by Western blotting,and the mRNA expressions of NF-κB subunit p65 and set7/9 were determined by real timequantitative PCR.The expression of monocyte chemoattractant protein 1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) were detected by enzyme-linked immunosorbent assay.Results (1)Compared with those in normal control group,the expressions of H3K4mel protein and set7/9 mRNA were first up-regulated in high glucose group,then gradually down-regulated in the following 48 h normal-glucose medium (as compared with those at 0 h,all P ＜ 0.05).At 72 h there was no statistic difference between high glucose group and normal control group (all P ＞ 0.05).(2) Compared with those in normal control group,the up-regulated p65 mRNA,VCAM-1 and MCP-1 sustained at least for 72 h in high glucose group.Conclusions Transient high glucose can induce persistent inflammatory factors expression in rat glomerular mesangial cells,which may via histone modification.

Objective To analyze the risk factors and correlation between clinical indicators and the four main pathological lesions of IgA ne?phropathy in the Oxford classification:mesangial hypercellularity(M0/1),endocapillary proliferation(E0/1),segmental sclerosis or adhesion(S0/1), and tubular atrophy/interstitial fibrosis(T0/1/2). Methods Clinical and pathological data were collected from 514 patients with biopsy?proven IgA nephropathy admitted in our hospital from February 17,2006 to October 11,2011. These patients were all above 18 years old. Cases with sec?ondary causes of mesangial IgA deposition were excluded,such as Henoch?chonlein purpura,ankylosing spondylitis and psoriasis et al. The inde?pendent risk factors affecting the pathological classification were analyzed by Spearman rank correlation analysis and two?category and multi?classi?fication logistic regression using SPSS 17.0 statistical software. Results In 514 IgAN patients,the ratio of males to females was 1.06:1. The aver?age age was 35.70±11.99 years,and the average disease duration was 18.31±30.42 months. M0E0S0T0 was the major pathologic classification of isolated hematuria. Chronic kidney disease(CKD)stage,24 hours proteinuria,albuminuria,urine transferrin and IgG levels were positively corre? lated with M lesion;serum albumin,C3 and PLT showed a negative correlation with M lesion. Twenty four hours proteinuria and blood platelet count were the independent risk factors for M lesion. As shown by stratified analysis ,the proportion of M1 in cases with 24 hours proteinuria≥3.5 g/d is much higher than that in cases with non?nephrotic range proteinuria. Age,systolic blood pressure,uRBC,24 hours proteinuria,albuminuria urine transferrin and IgG levels were positively correlated with E lesion,Duration,serum albumin showed a negative correlation with E lesion. Age and duration of nephritis were independent risk factors for E lesion. 73.3%of patients that above 60 years old showed endothelial proliferation. CKD stage,24 hours proteinuria were positively correlated with S lesion. Age,CKD stage,systolic blood pressure,diastolic blood pressure,C4,TC, LDL?C,CRP,Fib,UA,Cys?C and 24 hours proteinuria,urineβ2?microglobulin,albumin,transferrin and IgG levels were positively associated with T lesion;hemoglobin,serum albumin,serum IgG showed a negative correlation with T lesion. Infection history,high CRP levels,DBP more than 90 mmHg,hypoalbuminemia,high low density lipoproteinemia,and anemia were independent risk factors for T lesion. Conclusion Twenty four hours proteinuria,blood platelet count,age,duration of nephritis,hypoalbuminemia,anemia,hyperlipidemia,DBP≥90 mmHg and high CRP lev?els were risk factors for the Oxford classification of IgA nephropathy. Renal biopsy should be carried out in time to make clear the pathological clas?sification and individual treatment,so as to improve the prognosis.

Objective To elucidate the efficiency lncRNA GAS5 and miR-21 as biomarkers in diabetes mellitus and diabetic nephropathy.Methods The patients were divided into three groups,diabetic nephropathy group (DN group proven by renal biopsy,n=25,14 males and 11 females),diabetes group (DM group,with normal urine albumin creatinine ratio,n=10,4 males and 6 females),and normal control group (NC group,n=9,4 males and 5 females).The expressions of lncRNA GAS5 and miR-21 in serum samples were detected by real-time quantitative PCR.The correlation between serum lncRNA GAS5 and miR-21 expressions and the clinical parameters was analyzed by T-test,Pearson,Spearman test and multivariate linear regression analysis.Differences of lncRNA GAS5 and miR-21 in different groups were analyzed by one-way analysis of variance.The ROC curve was used to analyze the diagnostic efficacy of lncRNA GAS5 and miR-21 in diabetes and diabetic nephropathy.All data were analyzed by SPSS 20.0 and GraphPad software,with P ＜ 0.05 as considered statistically significant.Results (1) The expression of serum lncRNA GAS5 was significantly down-regulated and serum miR-21 was significantly up-regulated in both diabetes mellitus and diabetic nephropathy patients compared to the NC group all (P ＜ 0.05).(2) In DN patients,the expression of serum lncRNA GAS5 was gradually up-regulated along with the increment of 24 h urinary protein.The expression of serum miR-21 was gradually up-regulated along with renal biopsy stage Ⅱb-Ⅲ of DN (P ＜ 0.05).(3)FBG and HbA1c were all negatively correlated with serum lncRNA GAS5 (P ＜ 0.05),and FBG was independently correlated with serum lncRNA GAS5 (P ＜ 0.05).Urine microalbumin,Total cholesterol (TC),Scr,Urea and SBP were all positively correlated with serum miR-21(P ＜ 0.05).Albumin (ALB)and estimated GFR (eGFR) were negatively correlated with serum miR-21(P ＜ 0.05),and ALB was independently correlated with serum miR-21 (P ＜ 0.05).(4) The diagnostic efficiency of serum lncRNA GAS5,miR-21 and lncRNA GAS5/miR-21 as "diagnostic signature" for DM were was good (P ＜ 0.05).(5) The diagnostic efficiency of serum miR-21 and lncRNA GAS5/miR-21 as "diagnostic signature" for DN were was good (P ＜ 0.05).Conclusions (1) Serum lncRNA GAS5 had good diagnostic efficiency in diabetes mellitus.The sensitivity of lncRNA GAS5/miR-21 for diagnosis of diabetes was 85.71％,and specificity was 88.89％.(2) The level of serum miR-21 can be used as a noninvasive diagnostic marker for diabetic nephropathy.

Objective To observe the expression of microRNA-148b (miR-148b) induced by high glucose in rat mesangial cells,and to explore its effect on its target gene AMP-activated protein kinase α1 (AMPKα1) and extracellular matrix excretion.Methods Rat mesangial cells were divided ino 3 groups:normal glucose (NG,5.5 mmol/L glucose) group,hypertonic (MA,5.5 mmol/L glucose+19.5 mmol/L mannitol) group and high-glucose (HG,25.0 mmol/L glucose) group.MiR-148b expression was detected by real time PCR.Then miR-148b inhibitor was transfected to rat mesangial cells.Their protein expressions of AMPKα1,glucose regulated protein 78 (GRP78),C/EBP homologous protein (CHOP),fibronectin (FN) and collagen Ⅳ were detected by Western blotting.The expression of AMPKα1 mRNA was detected by real time PCR.The expression of collagen Ⅳ was also detected by immunofluorescence.Results Compared with NG group,HG group showed up-regulated miR-148bexpression,down-regulated AMPKαl mRNA and protein expressions,and up-regulated CHOP,GRP78,collagen Ⅳ and FN expressions (all P ＜ 0.05).HG-induced mesangial cells with miR-148binhibitor had up-regulated AMPKα1 mRNA and protein expressions,and down-regulated CHOP,GRP78,collagen Ⅳ,FN expressions as compared with HG-induced cells without miR-148b inhibitor (all P ＜ 0.05).Conclusions HG can up-regulate miR-148b expression and down-regulate AMPKα1 expression in rat mesangial cells,then activate endoplasmic reticulum stress to induce extracellular matrix excretion.MiR-148b inhibitor up-regulates AMPKα1 expression,inhibits endoplasmic reticulum stress and reduces extracellular matrix excretion.