Regulatory factors of Nrf2 in age-related macular degeneration pathogenesis

Age-related macular degeneration(AMD)is a complicated disease that causes irreversible visual impairment.Increasing evidences pointed retinal pigment epithelia(RPE)cells as the decisive cell involved in the progress of AMD,and the function of anti-oxidant capacity of PRE plays a fundamental physiological role.Nuclear factor erythroid 2 related factor 2(Nrf2)is a significant transcription factor in the cellular anti-oxidant system as it regulates the expression of multiple anti-oxidative genes.Its functions of protecting RPE cells against oxidative stress(OS)and ensuing physiological changes,including inflammation,mitochondrial damage and autophagy dysregulation,have already been elucidated.Understanding the roles of upstream regulators of Nrf2 could provide further insight to the OS-mediated AMD pathogenesis.For the first time,this review summarized the reported upstream regulators of Nrf2 in AMD pathogenesis,including proteins and miRNAs,and their underlying molecular mechanisms,which may help to find potential targets via regulating the Nrf2 pathway in the future research and further discuss the existing Nrf2 regulators proved to be beneficial in preventing AMD.

Relationship between Apoptosis and E-cadherin Expression in Bronchial Epithelium of Smoking Mouse

To investigate whether apoptosis is associated with cell adhesion in bronchial epithelium, and whether it contributes to the kinetics of injury and repair of surface epithelia, this study was performed for E-cadherin expression by using immunohistochemistry technique and for apoptosis by TUNEL method An animal model of smoking was used for this study The results showed that epithelial cells with membrane anchored E-cadherin decreased remarkably at several time points during 6 months of exposure to smoke ( P< 0 01) and then restored to normal level This fluctuation was associated exclusively with the alteration in number of apoptotic cells ( P <0 01) There was no significant difference in activation of nuclear transcription factor NF-kappa B among groups ( P> 0 05) All these suggested that apoptosis is associated with E-cadherin expression in bronchial epithelium of smoking mouse

Vitamin D differentially regulates Salmonella-induced intestine epithelial autophagy and interleukin-1β expression

AIM To investigate the effects of active vitamin D3 on autophagy and interleukin(IL)-1β expression in Salmonella-infected intestinal epithelial cells(IECs).METHODS Caco-2 cells, NOD2 siR NA-, Atg16L1 siR NA- or vitamin D receptor(VDR) siR NA-transfected Caco-2 cells were pretreated with 1,25-dihydroxyvitamin D3(1,25D3), and then infected by wild-type S. typhimurium strain SL1344. The conversion of LC3-I to LC3-II was detected by Western blot analysis and LC3+ autophagosome was analyzed by immunofluorescence. Caco-2 cells or VDR si RNA-transfected cells were pretreated with 1,25D3, and then infected by SL1344. Membrane protein and total RNA were analyzed by Western blot and RT-PCR for VDR and Atg16L1 protein and m RNA expression, respectively. Atg16L1 si RNA-transfected Caco-2 cells were pretreated by 1,25D3 and then infected with SL1344. Total RNA was analyzed by RT-PCR for IL-1β mR NA expression.RESULTS The active form of vitamin D, 1,25D3, showed enhanced VDR-mediated Atg16L1 mR NA expression, membranous Atg16L1 protein expression leading to autophagic LC3 II proteins expression and LC3 punctae in Salmonella-infected Caco-2 cells which was counteracted by Atg16L1 and VDR si RNA, but Atg16L1 mediated suppression of IL-1β expression. Thus, active vitamin D may enhance autophagy but suppress inflammatory IL-1β expression in Salmonella-infected IECs.CONCLUSION Active vitamin D might enhance autophagic clearance of Salmonella infection, while modulation of inflammatory responses prevents the host from detrimental effects of overwhelming inflammation.

Lipopolysaccharide triggers nuclear import of Lpcat1 to regulate inducible gene expression in lung epithelia

AIM:To report that Lpcat1 plays an important role in regulating lipopolysaccharide (LPS) inducible gene tran-scription. METHODS:Gene expression in Murine Lung Epithelial MLE-12 cells with LPS treatment or Haemophilus influenza and Escherichia coli infection was analyzed by employing quantitative Reverse Transcription Polymerase Chain Reaction techniques. Nucleofection was used to deliver Lenti-viral system to express or knock down Lpcat1 in MLE cells. Subcellular protein fractionation and Western blotting were utilized to study Lpcat1 nuclear relocation. RESULTS:Lpcat1 translocates into the nucleus from thecytoplasm in murine lung epithelia (MLE) after LPS treatment. Haemophilus influenza and Escherichia coli , two LPS-containing pathogens that cause pneumonia, triggered Lpcat1 nuclear translocation from the cytoplasm. The LPS inducible gene expression profile was determined by quantitative reverse transcription polymerase chain reaction after silencing Lpcat1 or overexpression of the enzyme in MLE cells. We detected that 17 out of a total 38 screened genes were upregulated, 14 genes were suppressed, and 7 genes remained unchanged in LPS treated cells in comparison to controls. Knockdown of Lpcat1 by shRNA dramatically changed the spectrum of the LPS inducible gene transcription, as 18 genes out of 38 genes were upregulated, of which 20 genes were suppressed or unchanged. Notably, in Lpcat1 overex-pressed cells, 25 genes out of 38 genes were reduced in the setting of LPS treatment.CONCLUSION:These observations suggest that Lpcat1 relocates into the nucleus in response to bacterial infection to differentially regulate gene transcriptional repression.

Advanced delivery strategies facilitating oral absorption of heparins

Heparins show great anticoagulant effect with few side effects,and are administered by subcutaneous or intravenous route in clinics.To improve patient compliance,oral administration is an alternative route.Nonetheless,oral administration of heparins still faces enormous challenges due to the multiple obstacles.This review briefly analyzes a series of barriers ranging from poorly physicochemical properties of heparins,to harsh biological barriers including gastrointestinal degradation and pre-systemic metabolism.Moreover,several approaches have been developed to overcome these obstacles,such as improving stability of heparins in the gastrointestinal tract,enhancing the intestinal epithelia permeability and facilitating lymphatic delivery of heparins.Overall,this review aims to provide insights concerning advanced delivery strategies facilitating oral absorption of heparins.

Assessment of Nutritional Status of Clinical Patients by Determining Normal Range of Oral Mucosal Apoptosis and Proliferation Rate

The normal range of oral mucosal cell apoptosis and proliferation rate through a larger sample of non-malnourished crowd was investigated,and the nutritional status of clinical patients was assessed.Of 194 clinical patients selected according to 'NRS2002' guidance,there were 167 non-malnourished patients and 27 malnourished cases,respectively.Twelve patients with toxic reactions of grade III after postoperative chemotherapy (POC) were chosen.The oral mucosal epithelial apoptosis and proliferation rate were measured by using flow cytometry.The statistical significance was processed by using unpaired t-test.The results showed that there was no significant difference in gender,age and body weight between malnourished and non-malnourished groups.The normal range of oral mucosal epithelial apoptosis and the proliferation rate was (27.50±1.50)% and (15.12±1.68)% in non-malnourished patients,and that was (19.90±4.14)% and (6.66±5.83)% in the malnourished patients,respectively.It is concluded that the normal range of oral mucosa cell apoptosis and proliferation rate is achieved,which can not be influenced by gender,age,weight and other factors,and could be used as a sensitive and accurate index to assess the nutritional status of clinical patients.

Effect of shRNA Inhibiting HIF1α Gene on TIMP1 Expression in RPE Cells

Small hairpin RNA （shRNA） was used to silence the HIF1α gene in human retinal pigment epithelial cells （RPE） under hypoxia in order to observe the effect of gene silencing on the expression of matrix metalloproteinase tissue inhibitor 1 （TIMP1）. By using chemical hypoxic inducer CoCl2 to mimic RPE hypoxic environment, shRNA against the targeting region of HIF1α mRNA sequence was synthesized by a method of in vitro transcription, and the HIF1α was interfered in RPE cultured under hypoxia （induced by 150 μmol/L CoCl2 ）. RT-PCR was employed to detect the expression of HIF1α and TIMP1. The expression levels of HIF1α and TIMP1 were measured by using Western blotting. The results showed that after the RPE were transfected with specific shRNA against HIF1α mRNA, RT PCR revealed that under hypoxia, the efficacy of HIF1α gene silencing in RPE was 83.4 %. Western blotting revealed that the expression levels of HIF1α protein was dramatically dropped. In addition, RT-PCR results demonstrated that the expression of TIMP1 mRNA was decreased by 28.9 %, and the expression levels of TIMP1 protein were also significantly reduced by Western blotting. It was suggested that shRNA targeted against HIF1α mRNA could effectively silence the HIF1α gene, subsequently effectively inhibit the hypoxia-induced up-regulation of TIMP1.

Poly(ADP-ribose),adherens junctions,vinculin and the actin cytoskeleton:Current evidence,future perspectives and implications

Poly(ADP-ribose)(PAR)is a highly negatively charged polymer.PAR is synthesized by poly(ADP-ribose)polymerases(PARPs)and is involved in the assembly and stabilization of macromolecular complexes.Here,the presence and putative roles of poly(ADP-ribosyl)ation(PARylation)associated to adherens junctions(AJ)and the actin cytoskeleton in epithelial and Schwann cells,is reviewed.The hypothesis generated by analogy,stating that PAR is associated to AJ in other cell types,is postulated.According to this hypothesis,PAR associated to puncta adherentia in chemical synapses would participate in plasticity,learning and memory.In turn,PAR associated to fascia adherens in cardiomyocytes,would affect heart beating.PARP inhibitors are currently under development and clinical testing.Basic research in different tissues will probably influence their clinical uses.

OBSERVATION ON REGIONAL DIFFERENCES OF SOME DEHYDROGENASES AND HYDROLASES IN RAT EPIDIDYMIS

The epididymal epithelia, by secretion, fluid reabsorption and transition, provide a favorable environment for sperm maturation. We observed, with histochemical method, the regional differences of four hydrolases and five dehydrogenases in caput, corpus and cauda of rat epididymis

The Effect of Sodium Nitrite on Induction of Apoptosis in Human Gastric Adenocarcinoma Epithelia (AGS) Cells

To examine the cytocidal effect of sodium nitrite on the cancer cell, we subjected human gastric adenocarcinoma epithelia (AGS) cells to various experimentation following exposure to sodium nitrite, and measured the resulting changes in the levels of cell death, lactate dehydrogenase (LDH) release, and caspase-3, -6, -8, and -9 activities. Our data revealed that, in AGS cells, treatment with ≥6.25 mM sodium nitrite for 8 h resulted in an obvious increase in cell death. LDH release was also markedly increased following sodium nitrite treatment, but at a concentration of ≥6.25 mM for 24 h. This increasing trend showed a positive correlation (r = 0.9564, P < 0.05). In addition, we detected pronounced increases in caspase activities with various concentrations of sodium nitrite: caspase-3 at ≥25 mM for 1 h, ≥12.5 mM for 3 h and 6 h;caspase-9 at 50 mM for 1 h and 3 h, and ≥6.25 mM for 6 h;and caspase-6 at 50 mM for 1 h and 3 h. We did not however, detect any observable increase in the activity of caspase-8 following sodium nitrite treatment at any concentration or for any duration of treatment in this study. This data demonstrates that, in AGS cells, higher concentrations or longer durations of treatment with sodium nitrite could exhibit a cytocidal effect, and that sodium nitrite could induce apoptosis via activation of the caspase-9, caspase-3 cascade (intrinsic pathway) and caspase-6.

Ablation of NTPDase2+ cells inhibits the formation of filiform papillae in tongue tip

Background: Lingual epithelia in the tongue tip are among the most rapidly regenerating tissues, but the mechanism of cell genesis in this tissue is still unknown. Previous study has suggested the existence of multiple stem cell pools in lingual epithelia and papillae. Like K14+ and Sox2+ cells, NTPDase2+ cells have characteristics of stem cells.Methods: We employed a system using doxycycline to conditionally ablate NTPDase2+ cells in lingual epithelia and papillae by regulated expression of the diphtheria toxin A(DTA) gene. Transgenic lines, which expressed the rtTA gene in NTPDase2+ cells, were produced by pronuclear injection of zygotes from C57 BL/6 mice using the BAC clone RP23-47 P18. The NTPDase2-rtTA transgenic mice were crossed with the TetO-DTA transgenic animals. The double transgenic mice were treated with doxycycline. Doxycycline(Dox) was diluted in 5% sucrose in water to a final concentration of 0.3-0.5 mg/mL and supplied as drinking water.Results: After 15 days of Dox induction, the expression of NTPDase2, Sox2 and K14 was ablated from lingual epithelia. DTA expression in NTPDase2+cells did not inhibit the turnover of GNAT3+ or PLCb2+ cells in taste buds,nor the expression of S100 b beneath lingual epithelia and papillae. After35 days ablation of NTPDase2+ cells, the basic structure of lingual epithelia and papillae remained intact. However, the ratio of cell to total tissue area was decreased in lingual epithelia and circumvallate(CV) papillae. DTA expression also inhibited the regeneration of filiform papillae on the dorsal surface of the tongue tip.Conclusions: These studies provide important insights into the understanding of dynamic equilibrium among the multiple stem cell populations present in the lingual epithelia and papillae.

Respiratory failure

950301 Endotoxin induced acute lung injury and theprotective effects with prostaglandin E1 in rabbits.WANG Jianxin(汪建新),et al.General Hosp,PLA,Beijing,100853.Med J Chin PLA 1995;20(1):36-38.A model of acute lung injury was successfully repro-duced by intravenous injection of E coli endotoxin(700μg/kg) to rabbits.It has been found that therewas a series of changes in the lungs in group B,such asgranulocyte seguestration,disturbance in

Illustrative Cases

A-1.7 days after an electric burn.A-2.40 days after treatment with MEBT.Thewound was filled with the regenerated tis-Sue.30 days after,the epithelia of thewound edge covered the wound to achievethe healing.B-1.A burn involving92 per cent body surfacea rea.B-2.B-3.30 days after treatment with MEBT,thewound was healed.(in zhanjiang)

The abnormal expression of E-cadherin in intrahepatic bile duct epithelia cells in biliary atresia and its relationship with apoptosis

To explore the relationship between the expression of E-cadherin and the apoptosis in intrahepatic bile duct epithelial cells in biliary atresia (BA).Methods The E-cadherin expression was demonstrated by immunohistochemical staining for the liver specimens from 38 children with BA and 16 normal children.The apoptotic intrahepatic bile duct epithelial cells in these specimens were visualized by TdT-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay,and the apoptotic index (AI) was calculated from the percentage of apoptotic cells in total cells.Results The intensity of E-cadherin expression in bile duct epithelial cells in BA group was lower than that in the normal control group (0.33±0.12 vs 0.62±0.20,P<0.01).On the other hand,the AI in BA group was significant higher than that in control group (51.74±19.93 vs 12.34±19.32,P<0.01).An inverse correlation was detected between the intensity of E-cadherin and the AI in the liver from children with BA.Conclusion The abnormal decrease of E-cadherin may lead to an increase of the apoptosis of intrahepatic bile epithelial cells in BA,resulting in developmental disorder of intrahepatic bile duct and ductal plate malformation in the liver.12 refs,4 figs,1 tab.

Dynamic RNA profiles in the small intestinal epithelia of cats after Toxoplasma gondii infection

Background Felids are the only definitive hosts ofToxoplasma gondii. However, the biological features of the feline small intestine followingT. gondii infection are poorly understood. We investigated the changes in the expression of RNAs (including mRNAs, long non-coding RNAs and circular RNAs) in the small intestinal epithelia of cats followingT. gondii infection to improve our understanding of the life cycle ofT. gondii and cat responses toT. gondii infection.Methods Fifteen cats were randomly assigned to five groups, and the infection groups were inoculated with 600 tissue cysts of theT. gondii Pru strain by gavage. The small intestinal epithelia of cats were collected at 6, 10, 14, and 30 days post infection (DPI). Using high-throughput RNA sequencing (RNA-seq), we investigated the changes in RNA expression. The expression levels of differentially expressed (DE) genes and non-coding RNAs (ncRNAs) identified by RNA-seq were validated by quantitative reverse transcription PCR (qRT-PCR). Differential expression was determined using the DESeq R package.Results In total, 207 annotated lncRNAs, 20,552 novel lncRNAs, 3342 novel circRNAs and 19,409 mRNAs were identified. Among these, 70 to 344 DE mRNAs, lncRNAs and circRNAs were detected, and the post-cleavage binding sites between 725 ncRNAs and 2082 miRNAs were predicted. Using the co-location method, we predicted that a total of 235 lncRNAs target 1044 protein-coding genes, while the results of co-expression analysis revealed that 174 lncRNAs target 2097 mRNAs. Pathway enrichment analyses of the genes targeted by ncRNAs suggested that most ncRNAs were significantly enriched in immune or diseases-related pathways. NcRNA regulatory networks revealed that a single ncRNA could be directly or indirectly regulated by multiple genes or ncRNAs that could influence the immune response of cats. Co-expression analysis showed that 242 circRNAs, mainly involved in immune responses, were significantly associated withT. gondii infection. In contrast, 1352 protein coding RNAs, mainly involved in nucleic acid process/repair pathways or oocyte development pathways, were negatively associated withT. gondii infection.Conclusions This study is the first to reveal the expression profiles of circRNAs, lncRNAs and mRNAs in the cat small intestine followingT. gondii infection and will facilitate the elucidation of the role of ncRNAs in the pathogenesis ofT. gondii infection in its definitive host, thereby facilitating the development of novel intervention strategies againstT. gondii infection in humans and animals.

Emerging Therapeutic Strategies in The Fight Against Primary Biliary Cholangitis

The liver has a vital role in many metabolic and regulatory processes in the body.Primary biliary cholangitis(PBC),previously known as primary biliary cirrhosis,is a chronic cholestatic autoimmune disease of the intrahepatic bile ducts associated with loss of tolerance to mitochondrial antigens.At this time there is no definitive cure for PBC;however,ursodeoxycholic acid(UDCA)has been shown to reduce injury when administered as the first line of treatment.Additional therapeutics can be given concurrently or as an alternative to UDCA to manage the symptoms and further curb disease progression.Currently,a liver transplant is the only potentially curative option when the patient has developed end-stage liver disease or intractable pruritus.This review aims to delineate the pathogenesis of primary biliary cholangitis and shed light on current therapeutic strategies in the treatment of PBC.

Regulation of epithelial function, differentiation, and remodeling in the epididymis

The epididymis is a single convoluted tubule lined by a pseudostratified epithelium. Specialized epididymal epithelial cells, the so-called principal, basal, narrow, and clear cells, establish a unique luminal environment for the maturation and storage of spermatozoa. The epididymis is functionally and structurally divided into several segments and sub-segments that create regionally distinct luminal environments. This organ is immature at birth, and epithelial cells acquire their fully differentiated phenotype during an extended postnatal period, but the factors involved in this complex process remain incompletely characterized. In the adult epididymis, the establishment of an acidic luminal pH and low bicarbonate concentration in the epididymis contributes to preventing premature activation of spermatozoa during their maturation and storage. Clear cells are proton-secreting cells throughout the epididymis, but principal cells have distinct acid/base transport properties, depending on their localization within the epididymis. Basal cells are located in all epididymal segments, but they have a distinct morphology depending on the segment and species examined. How this structural plasticity of basal cells is regulated is discussed here. Also, the role of luminal factors and androgens in the regulation of epithelial cells is reviewed in relation to their respective localization in the proximal versus distal regions of the epididymis. Finally, we describe a novel role for CFTR in tubulogenesis and epithelial cell differentiation.

Mutations in foregut SOX2^+ cells induce efficient proliferation via CXCR2 pathway

Identification of the precise molecular pathways involved in oncogene-induced transformation may help us gain a better understanding of tumor initiation and promotion. Here, we demonstrate that SOX2^+ foregut epithelial cells are prone to oncogenic transformation upon mutagenic insults, such as Kras^G12D and p53 deletion. GFP-based lineage-tracing experiments indicate that SOX2^+ cells are the cells-of-origin of esophagus and stomach hyperplasia. Our observations indicate distinct roles for oncogenic KRAS mutation and P53 deletion. p53 homozygous deletion is required for the acquisition of an invasive potential, and Kras^G12D expression, but not p53 deletion, suffices for tumor formation. Global gene expression analysis reveals secreting factors upregulated in the hyperplasia induced by oncogenic KRAS and highlights a crucial role for the CXCR2 pathway in driving hyperplasia. Collectively, the array of genetic models presented here demonstrate that stratified epithelial cells are susceptible to oncogenic insults, which may lead to a better understanding of tumor initiation and aid in the design of new cancer therapeutics.

Protective Effect of Tangshen Formula on Interstitial Cells of Cajal in Colon of Diabetic Rats

Objective:To explore the effect of Tangshen Formula(TSF),a Chinese herbal medicine,on interstitial cells of Cajal(ICC)in the colon of diabetic rats.Methods:Fifty-four male Wistar rats were randomly divided into normal control(NC,n=14)and high-fat diet(HFD)groups(n=40).After 6 weeks,the rats in the HFD group were injected intraperitoneally streptozotocin once(30 mg/kg).Thirty rats with fasting blood glucose higher than 11.7 mmol/L were randomly divided into diabetes(DM)and TSF groups,15 rats in each group.Rats in the NC and DM groups were intragastrically administered with saline,and those in the TSF group were given with TSF(2.4 g/kg)once daily for 20 weeks.Expression levels of Bax,Bcl-2,and caspase-3 in colonic smooth muscle layer were measured by Western blotting and immunohistochemical staining.The number of ICC was determined by immunohistochemical staining.Immunofluorescence was used for analyzing the ratio of classically activated macrophages(M1)and alternatively activated macrophages(M2)to total macrophages.Electron microscopy was used to observe the epithelial ultrastructure and junctions.Results:TSF appeared to partially prevented loss of ICC in DM rats(P<0.05).Compared with the NC group,expression levels of Bcl-2,Bax,caspase-3,and TNF-αas well as the ratio of M1 to total macrophages increased in DM rats(all P<0.05),and the ratio of M2 to total macrophages decreased(P<0.05 or P<0.01).Compared with the DM group,TSF decreased the expression levels of abovementioned proteins and restore M2 to total macrophages ratio(P<0.05 or P<0.01).TSF appeared to attenuate the ultrastructural changes of epithelia and improve the tight and desmosome junctions between epithelia reduced in the DM rats.Conclusions:Reduced number of ICC in DM rats may be associated with damage of the intestinal barrier.The protective effects of TSF on ICC may be through repair of the epithelial junctions,which attenuates inflammation and inflammation-initiated apoptosis in colon of DM rats.

Regulation of inflammation by the REGγ-proteasome

With the support from the National Natural Science Foundation of China,the National Basic Research Program of China,and the Science&Technology Commission of Shanghai Municipality,the research teamled by Prof.Li Xiaotao(李晓涛)at the Shanghai Key Laboratory of Regulatory Biology,Institute of Biomedical Sciences,East China Normal University collaborated with Prof.Xiao Jianru at