MicroRNA-630 alleviates inflammatory reactions in rats with diabetic kidney disease by targeting toll-like receptor 4

BACKGROUND Diabetic kidney disease(DKD)is a major complication of diabetes mellitus.Renal tubular epithelial cell(TEC)damage,which is strongly associated with the inflammatory response and mesenchymal trans-differentiation,plays a significant role in DKD;However,the precise molecular mechanism is unknown.The recently identified microRNA-630(miR-630)has been hypothesized to be closely associated with cell migration,apoptosis,and autophagy.However,the association between miR-630 and DKD and the underlying mechanism remain unknown.AIM To investigate how miR-630 affects TEC injury and the inflammatory response in DKD rats.METHODS Streptozotocin was administered to six-week-old male rats to create a hypergly cemic diabetic model.In the second week of modeling,the rats were divided into control,DKD,negative control of lentivirus,and miR-630 overexpression groups.After 8 wk,urine and blood samples were collected for the kidney injury assays,and renal tissues were removed for further molecular assays.The target gene for miR-630 was predicted using bioinformatics,and the association between miR-630 and toll-like receptor 4(TLR4)was confirmed using in vitro investigations and double luciferase reporter gene assays.Overexpression of miR-630 in DKD rats led to changes in body weight,renal weight index,basic blood parameters and histopathological changes.RESULTS The expression level of miR-630 was reduced in the kidney tissue of rats with DKD(P<0.05).The miR-630 and TLR4 expressions in rat renal TECs(NRK-52E)were measured using quantitative reverse transcription polymerase chain reaction.The mRNA expression level of miR-630 was significantly lower in the high-glucose(HG)and HG+mimic negative control(NC)groups than in the normal glucose(NG)group(P<0.05).In contrast,the mRNA expression level of TLR4 was significantly higher in these groups(P<0.05).However,miR-630 mRNA expression increased and TLR4 mRNA expression significantly decreased in the HG+miR-630 mimic group than in the HG+mimic NC group(P<0.05).Furthermore,the levels of tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),and IL-6 were significantly higher in the HG and HG+mimic NC groups than in NG group(P<0.05).However,the levels of these cytokines were significantly lower in the HG+miR-630 mimic group than in the HG+mimic NC group(P<0.05).Notably,changes in protein expression were observed.The HG and HG+mimic NC groups showed a significant decrease in E-cadherin protein expression,whereas TLR4,α-smooth muscle actin(SMA),and collagen IV protein expression increased(P<0.05).Conversely,the HG+miR-630 mimic group exhibited a significant increase in E-cadherin protein expression and a notable decrease in TLR4,α-SMA,and collagen IV protein expression than in the HG+mimic NC group(P<0.05).The miR-630 targets TLR4 gene expression.In vivo experiments demonstrated that DKD rats treated with miR-630 agomir exhibited significantly higher miR-630 mRNA expression than DKD rats injected with agomir NC.Additionally,rats treated with miR-630 agomir showed significant reductions in urinary albumin,blood glucose,TLR4,and proinflammatory markers(TNF-α,IL-1β,and IL-6)expression levels(P<0.05).Moreover,these rats exhibited fewer kidney lesions and reduced infiltration of inflammatory cells.CONCLUSION MiR-630 may inhibit the inflammatory reaction of DKD by targeting TLR4,and has a protective effect on DKD.

Lactose-free milk powder can effectively relieve diarrhea symptoms in weaning SD rats and children

Diarrhea has become the leading cause of illness and death among infants and young children in developing countries.Clinically,patients with diarrhea showed damaged intestinal epithelial villi,usually accompanied by lactase deficiency.In this study,we evaluated the therapeutic effects of lactose-free milk powder on rats and children with diarrhea.Antibiotic-associated diarrhea(AAD)model was established by gavage with antibiotic mixture in SD rats,followed by administration of milk powder containing lactose or not.The results showed that lactose-free milk powder ameliorated A AD-related diarrhea symptoms,and accelerated the recovery from diarrhea.And 16S sequencing results indicated lactose-free milk powder contributed to increase theα-andβ-diversity of intestinal flora,and restore the intestinal microbiota disorder.In conclusion,our data demonstrate that lactose-free milk powder could alleviate diarrhea by restoring gut microbiota and intestinal barrier function.

Study of molecular mechanisms underlying the medicinal plant Tripterygium wilfordii-derived compound celastrol in treating diabetic nephropathy based on network pharmacology and molecular docking

Background:Diabetic nephropathy(DN)is a serious complication of diabetes with rising prevalence worldwide.We aimed to explore the anti-DN mechanisms of the compound celastrol derived from the medicinal plant Tripterygium wilfordii.Methods:Celastrol-related targets were obtained from Herbal Ingredients’Targets(HIT)and GeneCards databases.DN-related targets were retrieved from GeneCards,DisGeNET,and Therapeutic Targets Database(TTD).A Protein-protein interaction(PPI)network was established using the Search Tool for the Retrieval of Interacting Genes(STRING)database.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed using ClusterProfiler.The cytoHubba plugin was used to select the top 10 hub targets.Molecular docking was performed employing PyMOL and AutoDock software.Cell counting kit-8(CCK-8)and flow cytometry assays were used to detect the viability and apoptosis of NRK-52E cells,respectively.The mRNA expression levels of mitogen-activated protein kinase 3(MAPK3),tumor necrosis factor(TNF),and AKT serine/threonine kinase 1(AKT1)in NRK-52E cells were assessed using quantitative real-time polymerase chain reaction(qRT-PCR).Results:We obtained sixty-six overlapping targets of celastrol and DN.GO and KEGG analyses demonstrated that the core targets of celastrol and DN were mainly involved in the inflammatory and immune response,oxidative stress,advanced glycation end products(AGEs)and their receptors(RAGEs)(AGE-RAGE),TNF,interleukin 17(IL-17),and MAPK signaling pathways.Finally,based on the good binding activity with celastrol,MAPK3,TNF,and AKT1 were identified as the foremost targets of celastrol.We observed that celastrol enhanced the viability of high glucose(HG)-treated NRK-52E cells and inhibited apoptosis in the in vitro assays.Moreover,celastrol decreased the mRNA expression levels of MAPK3,TNF,and AKT1 in high glucose(HG)-treated NRK-52E cells.Conclusion:Celastrol may treat DN by targeting APK3,TNF,and AKT1 and regulating inflammatory responses and oxidative stress through the AGE-RAGE,TNF,IL-17,and MAPK signaling pathways.

Emerging roles of non-coding RNAs in gastric cancer:Pathogenesis and clinical implications

Gastric cancer is a leading cause of cancer-related deaths. However, the mechanisms underlying gastric carcinogenesis remain largely unclear. The association of non-coding RNAs(nc RNAs) with cancer has been widely studied during the past decade. In general, nc RNAs have been classified as small nc RNAs, including micro RNAs(mi RNAs), and long non-coding RNAs(lnc RNAs). Emerging evidence shows that mi RNAs and lnc RNAs play key roles in the formation and progression of many cancers. In this review, we focus on the regulation of mi RNAs and lnc RNAs in gastric cancer. mi RNAs and lnc RNAs appear to be involved in gastric tumor growth, invasion, and metastasis and in establishment of the gastric tumor microenvironment through various mechanisms. Furthermore, we also discuss the possibilities of establishing mi RNAs and lnc RNAs as potential biomarkers and therapeutic targets for gastric cancer. Taken together, we summarize the emerging roles of nc RNAs in gastric cancer development and their possible clinical significance.

Rebamipide suppresses diclofenac-induced intestinal permeability via mitochondrial protection in mice

AIM: To investigate the protective effect and mechanism of rebamipide on small intestinal permeability induced by diclofenac in mice. METHODS: Diclofenac (2.5 mg/kg) was administered once daily for 3 d orally. A control group received the vehicle by gavage. Rebamipide (100 mg/kg, 200 mg/kg, 400 mg/kg) was administered intragastrically once a day for 3 d 4 h after diclofenac administration. Intestinal permeability was evaluated by Evans blue and the FITC-dextran method. The ultrastructure of the mucosal barrier was evaluated by transmission electron microscopy (TEM). Mitochondrial function including mitochondrial swelling, mitochondrial membrane potential, mitochondrial nicotinamide adenine dinucleotide-reduced (NADH) levels, succinate dehydrogenase (SDH) and ATPase activities were measured. Small intestinal mucosa was collected for assessment of malondialdehyde (MDA) content and myeloperoxidase (MPO) activity. RESULTS: Compared with the control group, intestinal permeability was significantly increased in the diclofenac group, which was accompanied by broken tight junctions, and significant increases in MDA content and MPO activity. Rebamipide significantly reduced intestinal permeability, improved inter-cellular tight junctions, and was associated with decreases in intestinal MDA content and MPO activity. At the mitochondrial level, rebamipide increased SDH and ATPase activities, NADH level and decreased mitochondrial swelling. CONCLUSION: Increased intestinal permeability induced by diclofenac can be attenuated by rebamipide, which partially contributed to the protection of mitochondrial function.

Effect of Shenqiyixin prescription on myocardial energy metabolism in patients with chronic heart failure

Objective:To observe the heart function、MEE and serum FFA of Shenqiyixin prescription for heart qi(yang)deficiency with blood stasis and fluid retention type of chronic heart failure patients,to evaluate the clinical efficacy and safety.Methods:64 cases with chronic heart failure of heart qi(yang)deficiency with blood stasis and fluid retention were randomly divided into two groups,3 cases were excluded,31 cases in the control group and 30 cases in the treatment group.The control group was treated with western medicine,and the treatment group was treated with Shenqiyixin prescription additionally.The course of treatment was 3 weeks.Observe the changes of TCM syndrome score,6MWD,LVEF,cESS,MEE,serum NTproBNP,serum FFA and safety indexes of each group before and after treatment.Results:after treatment,the TCM Syndromes of each group were improved.The total effective rate of the treatment group was 93.33%,and the control group was 83.87%.The treatment group was more effective(P<0.05).After treatment,the TCM syndrome score,cESS,MEE,NTproBNP and FFA of each group were decreased,6MWD and LVEF were increased(P<0.05),and the treatment group was superior to the control group(P<0.05).Conclusion:Shenqiyixin prescription can improve the TCM syndrome and heart function in patients with chronic heart failure of heart qi(yang)deficiency with blood stasis and fluid retention,at the same time,it can reduce the level of MEE and serum FFA in patients with heart failure.

单纯型大疱性表皮松解症伴肌营养不良1例

单纯型大疱性表皮松解症伴肌营养不良(epidermolysis bullosa simplex with muscular dystrophy, EBS-MD)由PLEC基因变异引起, 主要特征为出生时或生后不久出现皮肤水疱及进行性肌无力。本文报道1例危重型EBS-MD患儿, 以提升儿科医生识别与诊治这类疾病的能力。

Low level of ARID1A contributes to adaptive immune resistance and sensitizes triple-negative breast cancer to immune checkpoint inhibitors

Background Immune checkpoint inhibitors(ICIs)shed new light on triple-negative breast cancer(TNBC),but only a minority of patients demonstrate response.Therefore,adaptive immune resistance(AIR)needs to be further defined to guide the development of ICI regimens.Methods Databases,including The Cancer Genome Atlas,Gene Ontology Resource,University of California Santa Cruz Genome Browser,and Pubmed,were used to screen epigenetic modulators,regulators for CD8+T cells,and transcriptional regulators of programmed cell death-ligand 1(PD-L1).Human peripheral blood mononuclear cell(Hu-PBMC)reconstruction mice were adopted for xenograft transplantation.Tumor specimens from a TNBC cohort and the clinical trial CTR20191353 were retrospectively analyzed.RNA-sequencing,Western blotting,qPCR and immunohistochemistry were used to assess gene expression.Coculture assays were performed to evaluate the regulation of TNBC cells on T cells.Chromatin immunoprecipitation and transposase-accessible chromatin sequencing were used to determine chromatin-binding and accessibility.Results The epigenetic modulator AT-rich interaction domain 1A(ARID1A)gene demonstrated the highest expression association with AIR relative to other epigenetic modulators in TNBC patients.Low ARID1A expression in TNBC,causing an immunosuppressive microenvironment,promoted AIR and inhibited CD8+T cell infiltration and activity through upregulating PD-L1.However,ARID1A did not directly regulate PD-L1 expression.We found that ARID1A directly bound the promoter of nucleophosmin 1(NPM1)and that low ARID1A expression increased NPM1 chromatin accessibility as well as gene expression,further activating PD-L1 transcription.In Hu-PBMC mice,atezolizumab demonstrated the potential to reverse ARID1A deficiency-induced AIR in TNBC by reducing tumor malignancy and activating anti-tumor immunity.In CTR20191353,ARID1A-low patients derived more benefit from pucotenlimab compared to ARID1A-high patients.Conclusions In AIR epigenetics,low ARID1A expression in TNBC contributed to AIR via the ARID1A/NPM1/PD-L1 axis,leading to poor outcome but sensitivity to ICI treatment.

Age-induced changes in lung microenvironment: from melanoma dormancy to outgrowth

In a recent study published in Nature,Mitchell E.Fane et al.revealed the critical role of the aged microenvironment in the reactivation of melanoma cells from dormancy.1 The authors found that age-induced lung fibroblast secretory changes promoted growth activation of dormant melanoma cells in the lung,and age-induced skin microenvironment changes suppressed melanoma growth but promoted melanoma cell dissemination(Fig.1).

Potential targeted therapy and diagnosis based on novel insight into growth factors,receptors,and downstream effectors in acute kidney injury and acute kidney injury-chronic kidney disease progression

Acute kidney injury(AKI)is defined as a rapid decline in renal function and is characterized by excessive renal inflammation and programmed death of resident cells.AKI shows high morbidity and mortality,and severe or repeated AKI can transition to chronic kidney disease(CKD)or even end-stage renal disease(ESRD);however,very few effective and specific therapies are available,except for supportive treatment.Growth factors,such as epidermal growth factor(EGF),insulin-like growth factor(IGF),and transforming growth factor-β(TGF-β),are significantly altered in AKI models and have been suggested to play critical roles in the repair process of AKI because of their roles in cell regeneration and renal repair.In recent years,a series of studies have shown evidence that growth factors,receptors,and downstream effectors may be highly involved in the mechanism of AKI and may function in the early stage of AKI in response to stimuli by regulating inflammation and programmed cell death.Moreover,certain growth factors or correlated proteins act as biomarkers for AKI due to their sensitivity and specificity.Furthermore,growth factors originating from mesenchymal stem cells(MSCs)via paracrine signaling or extracellular vesicles recruit leukocytes or repair intrinsic cells and may participate in AKI repair or the AKI-CKD transition.In addition,growth factor-modified MSCs show superior therapeutic potential compared to that of unmodified controls.In this review,we summarized the current therapeutic and diagnostic strategies targeting growth factors to treat AKI in clinical trials.We also evaluated the possibilities of other growth factorcorrelated molecules as therapeutic targets in the treatment of AKI and the AKI-CKD transition.

Driving role of macrophages in transition from acute kidney injury to chronic kidney disease

Acute kidney injury(AKI),characterized by acute renal dysfunction,is an increasingly common clinical problem and an important risk factor in the subsequent development of chronic kidney disease(CKD).Regardless of the initial insults,the progression of CKD after AKI involves multiple types of cells,including renal resident cells and immune cells such as macrophages.Recently,the involvements of macrophages in AKI-to-CKD transition have garnered significant attention.Furthermore,substantial progress has also been made in elucidating the pathophysiological functions of macrophages from the acute kidney to repair or fibrosis.In this review,we highlight current knowledge regarding the roles and mechanisms of macrophage activation and phenotypic polarization,and transdifferentiation in the development of AKI-to-CKD transition.In addition,the potential of macrophage-based therapy for preventing AKI-to-CKD transition is also discussed.

Temperature-regulated self-assembly of lipids at free bubbles interface:A green and simple method to prepare micro/nano bubbles

Micro/nanobubbles play an essential role in ultrasound-based biomedical applications.Here,a green and simple method to fabricate micro/nanobubbles was developed by the temperature-regulated self-assembly of lipids in the presence of free bubbles.The self-assembly mechanism of lipids interacting with gas-water interfaces was investigated,and the ultrasound imaging of the obtained lipid-encapsulated bubbles(LBs)was further confirmed.Above the phase transition temperature(Tm),fluid lipids transform from vesicles to micelles,and further assemble to the free bubbles interface to be a compressed monolayer,resulting in lipid shelled microbubbles.Cooling below 7m induces the lipid shell to glassy state and stables the LBs.Moreover,increasing the 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000](DSPE-PEG2K)content in lipids formulation can further manipulate the shell curvature and reduce the LBs size into nanobubbles.LBs with diameters of 1.68±0.11 pm,704±7 nm and 208±6 nm were successfully prepared.The in vitro and in vivo ultrasound imaging results showed that all of the LBs had excellent echogenicity.The nanosized LBs revealed elongated imaging duration time and greater microvascular details for the liver tissue.Avoiding the organic solvent and complicated multiple preparation process,this method has great potential in construction of various multifunctional micro/nanobubbles with size control for theranostic applications.

Protective effect of indomethacin in renal ischemia-reperfusion injury in mice

Objective： To evaluate the renoprotection effects of non-steroidal anti-inflammatory drugs （NSAIDs） in renal ischemia-reperfusion injury （IRI） and the cyclooxygenase （COX）-1/2 blockade association by indomethacin （IMT） in the mice model. Methods： After the left renal pedicle of mice was clamped, IMT was administrated by intraperitoneal injection with four doses： 1, 3, 5, and 7 mg/kg. Blood and kidney samples were collected 24 h after IRI. The renal functions were assayed by the cytokines and serum creatinine （SCr） using enzyme-linked immunosorbent assay （ELISA） kits. Kidney samples were analyzed by hematoxylin and eosin （H＆E） and immunohistochemistry stainings. Results： The mice administered with 5 mg/kg IMT had a marked reduction in SCr and significantly less tubular damage The tumor necrosis factor a （TNF-α） activity in renal homogenates and interleukin 6 （IL-6） activity in serum had a marked reduction at doses of 5 and 7 mg/kg IMT. The administration of 3 and 5 mg/kg IMT had a marked reduction in the ratio of thromboxane B2 to 6-keto-prostaglandin F1α. COX-1 and COX-2 stainings were weaker in 5 mg/kg IMT groups than that in the other groups. Conclusions： There was a dose response in the IMT function of renal IRI in mice, and IMT had a protective effect in a certain dose range. The effect of IMT on mice IRI was related to COX-1/2 blockades.

Drug delivery system based on nanobubbles

Ultrasound(US)medical technology is widely used due to its advantages of safety,speed,and low cost.With the rapid development of biomedical nanotechnology,micro/nanobubbles,as US contrast agents,are not only valuable in US imaging diagnosis but also show broad application prospects in drug delivery.Nanobubbles(NBs)have more advantages as drug delivery systems(DDSs)due to their smaller size and longer circulation time compared with microbubbles.In this review,the fundamentals of NBs,including classification,stability mechanism,and preparation methods are summarized.Then,the strategies of a DDS based on NBs,including direct delivery,codelivery,target delivery,and stimuli-responsive delivery are reviewed and discussed.The three main applications of NBs-based DDSs,namely tumor theranostics,brain drug delivery,and vascular disease theranostics were highlighted.The challenges and future directions of the NBs-based DDSs are also considered and discussed.

Lithology-controlled stress variations of Longmaxi shale–Example of an appraisal wellbore in the Changning area

The Longmaxi shale is an extensive,prolific unconventional play in southwestern China.Its development in the Changning area is affected by ineffective hydraulic fracturing(HF)stimulation,fault reactivation and casing damage.It is suspected that the stress contrast within and between the shale reservoirs and the formations above and below matters to hydraulic fracture propagation and reservoir stimulation.To this end,the Longmaxi shale in the Changning area deserves a dedicated quantification of the in situ stress state and its variations.In this study,we re-visit the available data from one of the play’s first appraisal wellbores(X01)for an integrated geomechanics study,focusing on profiling the stress across the Longmaxi and its adjacent formations.Combining geophysical logs and other stress indicators,we re-interpret its stress profile in the context of lithological variations.The resulting stress variations are modeled primarily through a viscoplastic stress relaxation framework,compared with the results via the frictional equilibrium and an elastic theory(the Extended Eaton model).We offer some discussions on the differences and similarities of these stress profiling methods,and examine their applicability to Longmaxi shale in the Changning area.Our objective is to connect the lithology-controlled stress variations to the first-order complexities(HF ineffectiveness and fault reactivation)that have been observed in the area to date.

Proteins induced by telomere dysfunction are associated with human IgA nephropathy

Aging is one of the contributing risk factors for kidney diseases. Accumulating evidence prompts the view that telomere length in kidney tissue cells is an indicator for organismal aging. Previously identified aging markers （cathelin-related antimicrobial peptide （CRAMP）, stathmin, elongation factor-1α （EF-1α）, and chitinase） were associated not only with telomere driven aging in mice but also with human aging and chronic diseases. This study focuses on the relationship between these biomarkers and IgA nephropathy （IgAN） progression in the Chinese population. For 260 individuals, the four markers are determined in blind datasets using direct enzyme-linked immunosorbent assay （ELISA） and immunofluorescence staining. The expression levels of CRAMP and chitinase increased in blood plasma, urine, and kidney tissues during human IgAN progression. And for the other nephropathy, such as systemic lupus erythematosus （SLE）, diabetic nephropathy （DN）, and focal segmental glomerulosclerosis （FSGS）, there is no protein upregulaUon with telomere shortening. Moreover, a combination of CRAMP and chitinase can distinguish patients with IgAN from healthy individuals with 88.2%/92.5% （plasma） and 74.3%/84.2% （urine） sensitivity/specificity. These data provide the experimental evidence that telomere shortening and related inflammatory proteins are associated with human IgAN, and it could be a new direction for the disease progression study.