Tankyrase 2 promotes lung cancer cell malignancy

BACKGROUND Tankyrase 2(TNKS2)is a potential candidate molecular target for the prognosis and treatment of non-small cell lung cancer(NSCLC),but its biological functions are unclear.AIM To investigate the biological functions of TNKS2 in NSCLC.METHODS Using a lentiviral vector,we generated H647 model cells with TNKS2 knockdown by RNA interference and A549 model cells with TNKS2 overexpression by tran-sfection with a TNKS2 overexpressing plasmid.Increased and decreased exp-ression levels of TNKS2 in the two cell lines were verified using real-time reverse transcriptase-polymerase chain reaction and Western blot analyses.Cell apopto-sis,proliferation,and migration were determined using flow cytometry,carbo-xyfluorescein succinimidyl ester staining,and scratch assay,respectively.Im-munofluorescence staining was conducted to examine TNKS2 andβ-catenin ex-pression levels in the two transfected cell lines and the non-transfected cells.RESULTS TNKS2 mRNA and protein expression was significantly higher in the highly malignant NCI-H647 cells,while it remained at a low level in the less malignant A549 cells.Lentivirus-mediated overexpression of TNKS2 in A549 cells resulted in a 3-fold increase in gene expression and a 1.7-fold increase in protein expression(P<0.01).Conversely,shRNA interference targeting TNKS2 Led to an 8-fold decrease in gene expression and a 3-fold decrease in protein expression(P<0.01)in NCI-H647 cells.Furthermore,the cell apoptosis rate was significantly reduced(50%)and cell migration rate was increased(35%)in the TNKS2 overexpression group than in the control group(P<0.05).In contrast,shTNKS2 promoted apoptosis by more than one fold and reduced migration by 60%(P<0.05).Immunofluorescence analysis revealed enhanced nuclear localization ofβ-catenin fluorescence signal associated with high TNKS2 expression levels.Western blot analysis investigating TNKS2/β-catenin-related proteins indicated consistent changes between TNKS2 andβ-catenin expression in lung cancer cells,whereas Axin displayed an opposite trend(P<0.05).CONCLUSION The obtained results revealed that TNKS2 may serve as an adverse prognostic factor and a potential therapeutic target in NSCLC.

Methylenetetrahydrofolate reductase C677T and A1298C polymorphisms and gastric cancer susceptibility

AIM: To identify the association between methylenetetrahydrofolate reductase (MTHFR) polymorphisms and gastric cancer (GC) susceptibility.

FABRICATION OF SELF-ASSEMBLY DNA-C_(60) MULTILAYER FILMS

Electrostatic layer-by-layer self-assembly multilayer films were successfully fabricated from C-60-ethylenediamine adduct (C-60-EDA) and DNA. Under visible light irradiation, DNA is ready to be cleaved and the films are destroyed.

Experimental study of miRNA-210 modified MSCs in the treatment of myocardial ischemia-reperfusion injury in rats

Objective:To investigate the therapeutic effect of microRNA210(miRNA-210)modified mesenchymal stem cells(MSCs)on myocardial ischemia-reperfusion injury(MIRI)model rats.Methods:One SD rat was sacrificed,and the lower extremity tibia and femur were isolated.MSCs were cultured by whole bone marrow adherence method to construct miRNA-210 modified MSCs.40 SD rats were divided into the sham operation group,model group,MSCs group,and miRNA-210+MSCs group,with 10 rats in each group.The left anterior descending coronary artery was ligated to prepare a model of myocardial ischemia and reperfusion.After successful modeling,50μl of MSCs suspension was injected into the tail vein of the MSCs group,and 50μl of miRNA-210 modified MSCs suspension was injected into the tail vein of the miRNA-210+MSCs group.The sham operation group and the model group were injected with the same amount of normal saline.On the 10th day after modeling,the area of myocardial infarction,morphological changes of myocardial tissue,myocardial cell apoptosis rate,and miRNA-210 expression were compared in each group.Results:The area of myocardial infarction and the rate of myocardial cell apoptosis in the model group were significantly higher than those in the sham operation group(<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the MSCs group were significantly lower than those in the sham operation group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly higher than those in the MSCs group(P<0.05);The area of myocardial infarction and the rate of myocardial cell apoptosis in the miRNA-210+MSCs group were significantly lower than those in the sham operation group(P<0.05).The expression level of miRNA-210 in the myocardial tissue of the model group was significantly higher than that in the sham operation group(P<0.05);There were no significantly different in the expression level of miRNA-210 in the myocardial tissue between the MSCs group and model group(P>0.05);The expression level of miRNA-210 in the myocardial tissue of MSCs group was significantly higher than in the MSCs group,model group and sham operation group(P<0.05).HE staining showed that the miRNA-210+MSCs group had normal morphology of myocardial tissues,more uniform cytoplasmic staining,and arranged neatly myocardial fibers.The inflammatory cell infiltration and interstitial edema of the miRNA-210+MSCs group were significantly improved compared with the model group and MSCs group.Conclusion:MiRNA-210 modified MSCs can inhibit myocardial cell apoptosis in myocardial ischemia-reperfusion injury model rats,reduce the area of myocardial infarction,and improve pathological damage of myocardial tissue in rats,which has a certain therapeutic effect on myocardial ischemia-reperfusion injury.

n-octyl acrylate is a candidate sex pheromone component involved in courtship in parasitoid wasp Microplitis mediator

Sex pheromones are considered to play critical roles in partner communication of most parasitic Hymenoptera.However,the identification of sex pheromone components remains limited to a few families of parasitoid wasps.In this study,we functionally characterized a candidate sex pheromone component in Microplitis mediator(Hymenoptera:Braconidae),a solitary parasitoid of Noctuidae insects.We found that the body surface extract from female wasps could significantly stimulate courtship behavior of males.Gas chromatography-electroantennographic detection(GC-EAD)analysis revealed that a candidate semiochemical from extract triggered significant electrophysiological response of antennae of males.By performing gas chromatography-mass spectrometer(GC-MS)measurement,GC-EAD active compound was identified as n-octyl acrylate,a candidate sex pheromone component in female M.mediator.In electroantennogram(EAG)tests,antennae of male wasps showed significantly higher electrophysiological responses to n-octyl acrylate than those of females.Y-tube olfactometer assays indicated that male wasps significantly chose n-octyl acrylate compared with the control.Furthermore,male wasps showed a remarkable preference for n-octyl acrylate in a simulated field condition behavioral trial;simultaneously,n-octyl acrylate standard could also trigger significant courtship behavior in males.We propose that n-octyl acrylate,as a candidate vital sex pheromone component,could be utilized to design behavioral regulators of M.mediator to implement the protection and utilization of natural enemies.

Glucose-sensitive Membrane with PBA-based Contraction-type Linear Polymer as Chemical Valves Prepared by Surface Grafting

Glucose-sensitive membrane has potential application in self-regulating insulin release.Phenylboronic acid(PBA)is a well-known glucose reporter.Unfortunately,most PBA-based glucose-sensitive materials are expansion-type,which are not suitable as chemical valves in membrane pores for self-regulating insulin release.According to a new glucose-sensitive mechanism,we synthesized PBA-based contraction-type glucose-sensitive liner polymer and microgels.Herein,a glucose-sensitive membrane was prepared by grafting PBA-based contraction-type glucose-sensitive linear polymer on the membrane surface.Through adjusting the chain length and chain density,the glucose-sensitivity of the membrane was optimized.The membrane can reversibly regulate insulin release at physiologically relevant glucose concentrations in simulates body fluids and fetal bovine serum.The membrane also has good stability,anti-fouling and biocom patibility.It has potential application in selfregulating insulin release.

Functional characterization of a Niemann-Pick type C2 protein in the parasitoid wasp Micropfitis mediator

Niemann-Pick type C2 （NPC2） is a type of small soluble protein involved in lipid metabolism and triglyceride accumulation in vertebrates and arthropods. Recent stud- ies have determined that NPC2 also participates in chemical communication of arthropods. In this work, two novel NPC2 proteins （MmedNPC2a and MmedNPC2b） in Microplitis mediator were identified. Real-time quantitative PCR （qPCR） analysis revealed that Mmed- NPC2a was expressed higher in the antennae than in other tissues of adult wasps compared with MmedNPC2b. Subsequent immunolocalization results demonstrated that NPC2a was located in the lymph cavities of sensilla placodea. To further explore the binding charac- terization of recombinant MmedNPC2a to 54 candidate odor molecules, a fluorescence binding assay was performed. It was found MmedNPC2a could not bind with selected fatty acids, such as linoleic acid, palmitic acid, stearic acid and octadecenoic acid. How- ever, seven cotton volatiles, 4-ethylbenzaldehyde, 3,4-dimethylbenzaldehyde, fl-ionone, linalool, m-xylene, benzaldehyde and trans-2-hexen-l-al showed certain binding abilities with MmedNPC2a. Moreover, the predicted 3D model of MmedNPC2a was composed of seven r-sheets and three pairs of disulfide bridges. In this model, the key binding residues for oleic acid in CjapNPC2 of Camponotus japonicus, Lue68, Lys69, Lys70, Phe97, Thr103 and Phe127, are replaced with Phe85, Ser86, His87, Leu113, Tyr119 and Ile143 in MmedNPC2a, respectively. We proposed that MmedNPC2a in M. mediator may play roles in perception of plant volatiles.

Application of next generation sequencing in the screening of monogenic diseases in China,2021:a consensus among Chinese newborn screening experts

Newborn screening(NBS)refers to a maternal and newborn healthcare technology,in which special examinations of congenital and genetic diseases that could seriously impact the health of newborns,are implemented during the neonatal period to provide early diagnosis and treatment[1].With a history of more than 60 years,NBS has advanced greatly due to technological progress resulting in significant improvement in the number of diseases covered by NBS and in screening efficiency[2-7].

Silencing of odorant binding protein gene AlinOBP4 by RNAi induces declining electrophysiological responses of Adelphocoris lineolatus to six semiochemicals

Odorant binding proteins （OBPs） are believed to be important for transporting semiochemicals through the aqueous sensillar lymph to the olfactory receptor cells within the insect antennal sensilla. Here, we injected AlinOBP4-siRNA into the conjunctivum between prothorax and mesothorax of the lucerne plant bug, Adelphocoris lineolatus and evaluated the silencing of AlinOBP4 by reverse transcription polymerase chain reaction （RT-PCR） analysis, quantitative real-time PCR （qPCR） test and electroantennogram （EAG） assay. The combination of RT-PCR and qPCR analyses revealed that the levels of mes- senger RNA transcript were significantly reduced ~95% in AlinOBP4-siRNA-treated A. lineolatus males and ~75% in RNAi-treated females within 48 hours. It was found that there are different EAG responses between male and female bugs when the AlinOBP4 gene was silenced by RNAi. The EAGs of A. lineolatus to two plant volatiles, tride- canal and hexyl alcohol, were reduced 9.09% and 79.45% in RNAi-treated males, 62.08% and 62.08% in RNAi-treated females compared to the controls, separately. Antennae of RNAi-treated bugs showed significantly lower electrophysiological responses to four sex pheromone analogs, butyl butanoate, 1-hexyl butyrate, （E）-2-hexenyl butyrate and hexyl hexanoate. The EAG recordings were reduced 35.43%, 35.24%, 39.96% and 78.47% in RNAi-treated males and 64.52%, 18.13%, 36.88% and 49.52% in RNAi-treated females, respectively. The results suggested that AlinOBP4 might play dual-roles in the identifi- cation of plant volatiles and sex pheromones. It was suspected that AlinOBP4 may have different functions in odor perception between male and female A. lineolatus.

Molecular characterization and expression of sensory neuron membrane proteins in the parasitoid Microplitis mediator (Hymenoptera: Braconidae)

Sensory neuron membrane proteins(SNMPs),homologs of the human fatty acid transport protein CD36 family,are observed to play a significant role in chemoreception,especially in detecting sex pheromone in Drosophila and some lepidopteran species.In the current study,two full‐length SNMP transcripts,MmedSNMP1 and MmedSNMP2,were identified in the parasitoid Microplitis mediator(Hymenoptera:Braconidae).Quantitative real‐time polymerase chain reaction analysis showed that the expression of MmedSNMP1 was significantly higher in antennae than in other tissues of both sexes.In addition,the MmedSNMP1 transcript was increased dramatically in newly emerged adults and there were no significant differences between adults with or without mating and parasitic experiences.However,compared with MmedSNMP1,the expression of MmedSNMP2 was widely found in various tissues,significantly increased at half‐pigmented pupae stage and remained at a relatively constant level during the following developmental stages.It was found that MmedSNMP1 contained eight exons and seven introns,which was highly conserved compared with other insect species.In situ hybridization assay demonstrated that MmedSNMP1 transcript was distributed widely in antennal flagella.Among selected chemosensory genes(odorant binding protein,odorant receptor,and ionotropic receptor genes),MmedSNMP1 only partially overlapped with MmedORco in olfactory sensory neurons of antennae.Subsequent immunolocalization results further indicated that MmedSNMP1 was mainly expressed in sensilla placodea of antennae and possibly involved in perceiving plant volatiles and sex pheromones.These findings lay a foundation for further investigating the roles of SNMPs in the chemosensation of parasitoids.

Gene cloning and expression of aminopeptidase N and cadherin from midgut of the rice stem borer, Chilo suppressalis

The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N （CsAPN） and a cadherin （CsCad） were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here （CsCad） showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region （CR7）, which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin CrylAb. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.

Characterization and target gene analysis of microRNAs in the antennae of the parasitoid wasp Microplitis mediator

MicroRNAs(miRNAs),a class of non-coding single-strand RNA molecules encoded by endogenous genes,are about 21–24 nucleotides long and are involved in the post-transcriptional regulation of gene expression in plants and animals.Generally,the types and quantities of miRNAs in the different tissues of an organism are diverse,and these divergences may be related to their specific functions.Here we have identified 296 known miRNAs and 46 novel miRNAs in the antennae of the parasitoid wasp Microplitis mediator by high-throughput sequencing.Thirty-three miRNAs were predicted to target olfactory-associated genes,including odorant binding proteins(OBPs),chemosensory proteins,odorant receptors(ORs),ionotropic receptors(IRs)and gustatory receptors.Among these,17 miRNAs were significantly highly expressed in the antennae,four miRNAs were highly expressed both in the antennae and head or wings,while the remaining 12 miRNAs were mainly expressed in the head,thorax,abdomen,legs and wings.Notably,miR-9a-5p and miR-2525-3p were highly expressed in male antennae,whereas miR-1000-5p and novel-miR-13 were enriched in female antennae.The 17 miRNAs highly expressed in antennae are likely to be associated with olfaction,and were predicted to target one OBP(targeted by miR-3751-3p),one IR(targeted by miR-7-5p)and 14 ORs(targeted by 15 miRNAs including miR-6-3p,miR-9a-5p,miR-9b-5p,miR-29-5p,miR-71-5p,miR-275-3p,miR-1000-5p,miR-1000-3p,miR-2525-3p,miR-6012-3p,miR-9719-3p,novel-miR-10,novel-miR-13,novel-miR-14 and novel-miR-28).These candidate olfactory-associated miRNAs are all likely to be involved in chemoreception through the regulation of chemosensory gene expression in the antennae of M.mediator.

Identification of heat shock cognate protein 70 gene （Alhsc70） of Apolygus lucorum and its expression in response to different temperature and pesticide stresses

Heat shock cognate protein 70 （Hsc70） is a very important stress-resistance protein of insects against environmental stresses. We employed fluorescent real-time quan- titative polymerase chain reaction and Western-blot techniques to analyze the transcrip- tional and translational expression profiles of AIHSC70 under extreme temperature （4℃ and 40℃） or 4 pesticide stresses in Apolygus lucorum. The results showed that the expres- sion of AIHSC70 were significantly induced by cyhalothrin or extremely high temperature （40℃） in both transcriptional and translational levels （P 〈 0.05）, while the transcriptional and translational level of AIHSC70 decreased significantly in treatments of chlorpyrifos or extreme cold temperature （4℃） （P 〈 0.05）. Moreover, afterApolygus lucorum treated by imidacloprid or emamectin benzoate, the expression of AIHSC70 was only up-regulated significantly at the transcriptional level （P 〈 0.05）, although obviously up-regulated at the translational level of AIHSC70. Therefore, this study confirmed that the Alhsc70 gene played important roles in response to both temperature and pesticide stresses, especially for cyhalothrin or extremely high temperature （40℃）. In addition, the significant polynomial regression correlations between temperature and the Alhsc70 expression level were shown in all the nymph and adult stages （P 〈 0.01）, indicating temperature was an important factor to affect the relative expression of Alhsc70.

Rice gene expression profiles responding to larval feeding of the striped stem borer at the 1st to 2nd instar stage

The objective of this study was to identify rice genes that are in response to the striped stem borer （SSB） （Chilo suppressalis Walker） feeding at the first to second larval stage. Using combined suppression subtractive hybridization （SSH） and dot blot approaches, we analyzed the induced defense genes that took place during the first 72 h of infesting intact rice （Oryza sativa L.） plants in sheath tissues with SSB larvae. By sequencing the whole SSH library, 39 expressed sequence tags involved in disease stress, insect stress or other stress responses were identified to be up-regulated by SSB larvae feeding. Among these genes, rice allene oxide cyclase （AOC）, terpene synthase （TPS） and four proteinase inhibitor （PI） genes were up-regulated by SSB larvae feeding. Real-time quantitative reverse transcription polymerase chain reaction analysis showed that four rice PI genes were already up-regulated at 6 h, and reached peaks between 6 h to 12 h. In addition, the transcription ofgene involving in jasmonate signaling pathway such as allene oxide cyclase （AOC） concerning rice early defense response to SSB feeding was activated after rice feeding by SSB for 2 h. Although the expression office terpene synthase （TPS） gene, involved in the biosynthesis ofmonoterpenes or diterpenes, was already up-regulated at 7 h, a significant increase in the expression was delayed until 12 h and reached its peak at 24 h. The present study identified six SSB-response genes and their expression patterns, which provides evidence and information to understand insect stress-response in plants.

Responsive Hybrid Poly(vinyl alcohol)Hydrogel Membranes with Embedded Microgels as Valves

The research and application of responsive materials have long been hampered by their complicated designs and tedious construction processes.Besides,many current responsive materials show retard or weak responsiveness.In this study,responsive hybrid poly(vinyl alcohol)hydrogel membranes with embedded poly(N-isopropylacrylamide-acrylic acid)microgels as valves were constructed by simple mixing and subsequent freezing-thawing process.In the structure of the membranes,the matrix poly(vinyl alcohol)chains thread through and entangle with the microgels,and the microgels are firmly constrained within the hybrid hydrogel network.The fast and sharp temperature responsiveness of the embedded microgels was largely retained and endowed the hydrogel membrane with excellent temperature and pH responsiveness.Moreover,the hydrogel membrane showed excellent fatigue resistance in both temperature and pH-responsive flux examination.This study presented the great potential of these hybrid hydrogel membranes in biomedical applications and provided a new strategy for the future design and construction of responsive biomaterials.

Associations between measures of pediatric human resources and the under-five mortality rate: a nationwide study in China in 2014

Background To quantify the associations between the under-five mortality rate(U5MR)and measures of pediatric human resources,including pediatricians per thousand children(PPTC)and the geographical distribution of pediatricians.Methods We analyzed data from a national survey in 2015-2016 in 2636 counties,accounting for 31 mainland provinces of China.We evaluated the associations between measures of pediatric human resources and the risk of a high U5MR(>18 deaths per 1000 live births)using logistic regression and restricted cubic spline regression models with adjustments for potential confounders.PPTC and pediatricians per 10,000 km^(2) were categorized into quartiles.The highest quartiles were used as reference.Results The median values of PPTC and pediatricians per 10,000 km^(2) were 0.35(0.20-0.70)and 150(50-500),respectively.Compared to the counties with the highest PPTC(≥0.7),those with the lowest PPTC(<0.2)had a 52% higher risk of a high U5MR,with an L-shaped relationship.An inverted J-shaped relationship was found that the risk of a high U5MR was 3.74[95%confidence interval(CI)2.55-5.48],3.07(95% CI 2.11-4.47),and 2.25 times(95% CI 1.52-3.31)higher in counties with<50,50-149,and 150-499 pediatricians per 10,000 km^(2),respectively,than in counties with≥500 physicians per 10,000 km^(2).The joint association analyses show a stronger association with the risk of a high U5MR in geographical pediatrician density than PPTC.Conclusion Both population and geographical pediatrician density should be considered when planning child health care services,even in areas with high numbers of PPTC.

Transgenic cotton expressing C YP392A4 double-stranded RNA decreases the reproductive ability of Tetranychus cinnabarinus

As a polyphagous pest, Tetranychus cinnabarinus has the ability to overcome the defense of various hosts, and causes severe losses to various economically important crops. Since the interaction between pest and host plants is a valuable clue to investigate potential ways for pest management, we intend to identify the key genes of T. cinnabarinus for its adaption on cotton, then, with RNA interference （RNAi） and transgenic technology, construct a transgenic cotton strain to interfere with this process, and evaluate the effect of this method on the management of the mites. The difference of gene expression of T. cinnabarinus was analyzed when it was transferred to a new host （from cowpea to cotton） through high-throughput sequencing, and a number of differentially expressed genes involved in detoxification, digestion and specific processes during the development were classified. From them, a P450 gene CYP392A4 with high abundance and prominent over-expression on the cotton was selected as a candidate. With transgenic technology, cotton plants expressing double-stranded RNA of CYP392A4 were constructed. Feeding experiments showed that it can decrease the expression of the target gene, result in the reduction of reproductive ability of the mites, and the population of T. cinnabarinus showed an apparent fitness cost on the transgenic cotton. These results provide a new approach to restrict the development of mite population on the host. It is also a useful attempt to control piercing sucking pests through RNAi and transgenic technology.

Epidemiology and region-specific risk factors for low Apgar scores in China:a nationwide study

Background Neonatal asphyxia is a serious public health issue.This study aimed to determine the epidemiology and region-specific risk factors for low Apgar scores,an important proxy for neonatal asphyxia,in China from 2015 to 2016.Methods The China Labor and Delivery Survey was a multicenter cross-sectional study including 96 hospitals distributed in 24(out of 34)provinces.Logistic regression analysis was performed to examine the risk factors for a low Apgar score(<7).Correspondence analyses were performed among neonates with low Apgar scores to explore the relationship between risk factors and geographical regions.The population attributable risk percentage(PAR%)was calculated for each region-specific risk factor.Results A total of 72,073 live births,including 320 births with low Apgar scores,were used for the analysis,giving a weighted rate of 3.9/1000 live births.There was a substantial difference in the incidence of low Apgar scores by geographic region,from 2.3/1000 live births in East China to 10.9/1000 live births in Northeast China.Maternal and obstetric factors are the major region-specific risk factors.In Southwest China,hypertensive disorders in pregnancy were more important contributors,with PAR% being 74.47%;in North and Northwest China,pre-pregnancy underweight was a more significant factor,with PAR% of 62.92%;in East China,infants born between 0:00 a.m.and 7:59 a.m.were a key factor,with PAR% of 80.44%.Conclusion Strategies based on region-specific risk factors should be considered to reduce the burden of low Apgar scores in China.