Objective To observe the effect of Danzhi Jiangtang Capsules(DJC) on serum interleukin-18 (IL-18),cystatin C (CysC) and relative biochemical indexes in patients with early diabetic kidney disease(DKD),and to explore its protective action on the kidney.Methods Sixty-two hospitalized patients with early diabetic nephropathy (period 11-Ⅲ) in Endocrinology Department of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine were divided into western medicine group and DJC group,31 cases in each group.The patients in western medicine group received conventional hypoglycemic drugs,and DJC group received DJC based on the treatment for the western medicine group.Before and after treatment,we observed the levels of serum IL-18,CysC,fasting plasma glucose (FPG),2-hour postprandial glucose (2hPG),glycosylated hemoglobin (HbA1c),serum creatinine (SCr),blood urea nitrogen (BUN),microalbumin (mAlb),urinary albumin /creatinine ratio (UACR),and estimated glomerular filtration rate (eGFR).Results The total effective rate was 90.0％ in DJC group and 76.7％ in the western medicine group,there being significant difference between the two groups(P ＜ 0.05).After treatment,the main traditional Chinese medical syndrome scores of DJC group were significantly lower than those before treatment,and the improvement was better than that of the western medicine group (P ＜ 0.05 or P ＜ 0.01).After treatment,the serum levels of IL-18,CysC,2hPG,SCr,BUN,mAlb,UACR in the two groups were significantly lower than those before treatment (P ＜ 0.05 or P ＜ 0.01),while eGFR level was higher than that before treatment (P ＜ 0.05 or P ＜ 0.01),and the improvement effect (except for BUN) in the DJC group was superior to that in the western medicine group(P ＜ 0.01).DJC had an effect on lowering the FPG and HbA1c (P ＜ 0.05 or P ＜ 0.01 compared with those before treatment),and the western medicine had no obvious effect on the two indexes (P ＞ 0.05).The results of Spearman correlation analysis showed that IL-18 was correlated with CysC,IL-18 and CysC were correlated with HbA1c,mALB and uACR.Conclusion DJC can protect renal function of patients with early diabetic nephropathy.

Objective To investigate the clinical roles of lysine methyltransferase 5A(KMT5A)in human hepatocellular carcinoma(HCC)and its functions in cell migration and invasion. Methods The expression levels of KMT5A of 60 cases were detected by immunohistochemistry(IHC). KMT5A siRNA was used to down-regulate the expression of KMT5A in SMMC-7721 cells. Cell migration and invasion were measured by wound healing assays and transwell assays,respectively. Immunoblotting was used to detect the expression of MMP-2 after siRNA trans-fection. miR-186 mimics were transfected into SMMC-7721 cells and the mRNA levels of KMT5A was detected by qRT-PCR after transfection. Results High expression of KMT5A was associated with large tumor diameter (>5 cm,P=0.047)and advanced TNM stage(Ⅲ+Ⅳ,P=0.035). The expression of KMT5A was knocked down by siRNA in SMMC-7721 cells. Down-regulation of KMT5A suppressed cell migration(P=0.031,P=0.006)and invasion(P=0.010),and impaired MMP-2 expression(P=0.040). Overexpression of miR-186 could significantly inhibit the expression of KMT5A(P = 0.007). Conclusions Over-expression of KMT5A in HCC tissues associ-ates with poor clinical features. KMT5A knockdown inhibits the migration and invasion on HCC cells.

Animals ; Humans ; Liver ; growth & development ; Liver Diseases ; Receptors, Notch ; metabolism ; Signal Transduction

Objective To investigate the effect of thyroid hormone on the expression of NF-κB and TNF-oα in the ischemic cortex of rats after focal cerebral ischemia-reperfusion.Methods 96 male SD rats were randomly divided into sham-operation group,sham-operation+T3 group,IR group and IR+T3 group.Using suture legal method to establish a rat model of middle cerebral artery occlusion for 2 h followed by reperfusion.In sham-operation+T3 group and IR+T3 group,T3 was given 3 days before ischemia and 1 hour after ischemia,respectively,intraperitoneal injection T3 10 μg/100 g for rats.Rats in other groups were given the same volume normal saline at the same time.The infarct size was determined by TTC staining at 24 h after reperfusion.HE staining was used to observe the morphological and structural changes of brain tissue.Using Real-time PCR method and immunohistochemical staining method to detect the expression of NF-κB mRNA,TNF-α mRNA and protein in ischemic cortex of rats.Results Compared with sham-operation group and sham-operation+T3 group,the pathological damage of brain tissue in IR group was obvious,while the pathologic damage of IR +T3 group was less than that in IR group.Immunohistochemistry assay showed that the expression of NF-κB was(49.19±5.55)in sham-operation group,(45.75±2.12) in sham-operation+T3 group,(56.88±2.23)in IR group and(50.25±1.67)in IR +T3 group,the expression of TNF-α was (22.50±3.07) in sham-operation group,(24.13±2.03) in sham-operation+T3 group,(37.25±2.82) in IR group and (30.25±1.67) in IR +T3 group,and the NF-κB,TNF-α in IR group were obviously higher than that in sham-operation group and sham-operation+T3 group(P＜0.05),while IR+T3 group were lower than that in IR group(P＜0.05).Real-time PCR showed that NF-κB mRNA,TNF-α mRNA level in IR group was the highest,which was higher than that of sham-operation group and sham-operation+T3 group(P＜0.05),and the NF-κB mRNA,TNF-oα mRNA expression in IR+T3 group were significantly decreased compared with that in IR group(P＜0.05).Conclusion Thyroid hormone has a protective effect on cerebral ischenia reperfusion injury,which may be achieved by reducing the expression of inflammatory factor NF-κB and TNF-oα.

Apoptosis is a process of programmed cell death that is controlled by genes. Normally, there are three regulation pathways involved in the process of apoptosis, including the signaling of intracel-lular mitochondria, endoplasmic reticula and extracellular death receptors. Recent studies showed that NF-κB is a key regulator in the process of apoptosis. NF-κB plays a promotional and a inhibitory role as well in the regulation of apoptosis, closely related to the the inhibitor of apoptosis proteins family, the B cell lymphoma/ lewkmia-2 family, tumor necrosis factor receptor associated factors, c-Jun N-terminal kinase, tumor necrosis factor related apoptosis inducing ligand and Fas-associated death domain protein-like interleukin-1β. Thus, investigation of the mechanism regarding NF-κB in apoptosis regulation is of great importance for apoptosis-related drug development. The paper reviews the recent research progress in the function of NF-κB in apoptosis pathway regulation.

This study chiefly aimed at investigating the effects of Dan Zhi Jiang Tang Capsule (DJC) on myocardial Toll-like receptor 4 (TLR4),tumor necrosis factor α (TNFα) and interleukin-8 (IL-8) and the cardiopathologic changes in diabetic rats.Fifty male rats,excluding 8 rats in the control group,had been administered with high fat forage for 4 weeks,which established the diabetic rat model combined with the intraperitoneal injection of streptozotocin (STZ) at 35 mg· kg-1.The established model rats were randomly divided into the model group,the DJC high-dose group,the DJC low-dose group and the pioglitazone group.The rats in the model group and the control group were intragastrically administered with DJC at 1.08 and 0.54 g· kg-1 each day,while those of the pioglitazone group were treated with pioglitazone at 10 mg· kg-1 each day,and those of the model group and control group with saline of the same volume.The administrations lasted 8 weeks with the continues high-fat diet.The levels of TLR4,TNFα and IL-8 protein in myocardium of the rats were determined,and so were the blood glucose,HbA1c and blood lipids.Compared with the control group,the expressions of myocardial TLR4,TNFα and IL-8 in the model group were significantly increased (P＜ 0.01).The expressions of TLR4,TNFα and IL-8 in the DJC high-dose and low-dose groups were significantly decreased (P＜0.05 or P＜0.01).The cardiopathological report showed severe myocardial lesion in the model group.However,myocardial lesion in the DJC high-dose group was significantly mitigated,which presented better therapeutic effects than the DJC low-dose group and the pioglitazone group.The levels of blood glucose,HbA1c,TG,TC,LDL-C in the model group were significantly higher than those in the control group (P＜0.05 or P＜0.01).The levels of blood glucose,HbA1c,TG,TC and LDL-C were significantly decreased in the DJC high-dose and low-dose groups in comparison with the model group (P＜ 0.05 or P＜ 0.01).In conclusion,DJC down-regulated the expressions of TLR4,TNFα and IL-8 and improved the degree of myocardial lesion in the diabetic rats,showing favorable hypoglycemic and lipid-lowering effects.

Objective To investigate the neuroprotective effect of thyroid hormones T3 on cerebral ischemia-reperfusion injury in rats and its mechanism.Methods SD rats were divided into four groups:sham+saline group,sham+T3 group,MCAO+saline group,MCAO+T3 group.The cerebral ischemia-reperfusion injury rat models were established by right middle cerebral artery occlusion.Thyroid hormones(10 μg/100 g)or normal saline were given respectively by intraperitoneal injection twice at 1 h after the onset of ischemia and 6 h after reperfusion.Neurobehavioral score was evaluated at 24 h after reperfusion;TTC staining was used to label infarction area;RT-PCR was used to detect the mRNA level of nerve growth factor(NGF)and brain derived neurotrophic factor(BDNF)in brain tissue;Western blot was employed to determine alterations in protein levels of NGF and BDNF.Results Compared with MCAO+saline group,the neurological deficit and the volume of cerebral infarction of MCAO+T3 group was decreased,and the mRNA and protein expression of NGF and BDNF of MCAO+T3 group were increased(P<0.05).Conclusion Thyroid Hormones could promote the nerve repair,stimulate the nerve regeneration and improve the nervous behavioral function by up-regulating the expression of NGF and BDNF.

OBJECTIVE To explore the effect and mechanisms of baicalein on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis in mice.METHODS BALB/c mice were randomly placed into three groups (n=10):normal control group,TNBS group,and TNBS+baicalein (20 mg· kg-1,once per day) group.Mouse colitis was induced by intrarectal injection of TNBS.Baicalein was administered by oral gavage two days prior to TNBS treatment and until the end of the study (a total of 9 d).The colon length was measured before HE staining was performed for histological damage assessment.The remaining colon pieces were collected to measure the content of tumor necrosis factor-α(TNF-α).Lipopolysaccharide (LPS)-stimulated RAW264.7 mouse macrophage was used as a cell model to determine the content of nitric oxide (NO) in cell culture medium,the mRNA levels of TNF-α,interleukin-6(IL-6),IL-1β,inducible nitric oxide synthase(iNOS),cyclooxygenase 2(COX-2) and monocyte chemoattractant protein-1 (MCP-1),and the protein expression of phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-κB (PI3K/AKT/NF-κB) pathway.RESULTS Baicalein significantly attenuated TNBS-induced colon shortening and histological injury (P＜0.05),which was correlated with the decline in the content of TNF-α in the colon.According to the jn vivo results,baicalein exposure down-regulated the secretion of NO and the mRNA expression of pro-inflammatory mediators (iNOS,COX-2,MCP-1,TNF-α,IL-1β and IL-6) in LPS-stimulated RAW264.7 cells (P＜0.05,P＜0.01).Additionally,the phosphorylation/activation of LPS-stimulated PI3K/AKT/NF-κB pathway was inhibited by baicalein treatment.CONCLUSION The beneficial effect of baicalein in TNBS-induced experimental colitis may be due to PI3K/AKT/NF-κB signaling inhibition.

Aim Toassesstheregulatoryeffectsofcar-damonin (CDN ) on toll-like receptor (TLR )-4/MyD88/NF-κB/iNOS signaling pathway in lipopolysac-charide (LPS )-stimulated RAW264. 7 macrophage cells.Methods LPS-stimulatedRAW264.7cells were divided into three groups:vehicle-treated group, LPS-treated group and LPS +CDN-treated group.Cell viability was assessed by CCK-8 assay.The concentra-tion of nitric oxide (NO)in cell culture medium was measured by Griess reagent.The mRNA levels of iN-OS,COX-2,MCP-1 ,TNF-α,IL-6 and IL-1βwere de-termined by reverse transcription real-time quantitative PCR(RT-qPCR).The protein levels of inducible nitric oxide synthase(iNOS),TLR4,myeloid differentiation factor 88(MyD88),nuclear factor κB(NF-κB)phos-phorylated (p )-p65 ,inhibitor κBα(IκBα),and p-IκBαweredeterminedbyWesternblot.Results 1~50 μmol·L-1 CDN had no cytotoxicity in RAW264. 7 cells.However,CDN inhibited the LPS-induced secre-tion of nitric oxide(NO)and mRNA expressions of iN-OS,COX-2,MCP-1 ,TNF-α,IL-6 and IL-1βin a dose-dependent manner.Moreover,50 μmol · L-1 CDN inhibited the LPS-induced up-regulation of iNOS, TLR4,MyD88,NF-κB p-p65,p-IκBαand down-reg-ulationofIκBα.Conclusion Cardamonininhibitsthe production of NO via a mechanism associated with the inhibition of TLR4/MyD88/NF-κB/iNOS pathway.

Aim To evaluate the effect and mecha-nism of vitexin in a mouse model of DSS-induced ulcer-ative colitis (UC).Methods C57BL/6 mice were randomly placed into three groups: normal control group,DSS group and DSS +Vitexin group.Mice coli-tis was induced by adding 4% dextran sulphate sodium (DSS)into the drinking water for seven days.Vitexin was administered once a day along with DSS treatment. Mice were monitored daily with body weight change and diarrhea symptoms.After sacrifice,colon was re-moved and fixed in 1 0% (W/V)buffered formalin for hematoxylin-eosin (H&E)staining.Histological dam-age was assessed as a combined score of inflammatory cell infiltration and mucosal damage.The remaining colon pieces were collected to measure the activity of myeloperoxidase (MPO)by ELISA method and to de-termine the mRNA expression of TNF-α,IL-6 and COX-2.Results None of the mice receiving vehicle alone exhibited body weight loss and mucosal disrup-tion at any point during the study.Vitexin treatment significantly ameliorated DSS-induced body weight loss and histological score.The activity of MPO and the mRNA expression of TNF-α,IL-6 and COX-2 were markedly inhibited by vitexin treatment.Conclusion Vitexin ameliorates DSS-induced colitis through sup-pressing leukocyte infiltration and pro-inflammatory mediators production.