Quantitative trait loci identification reveals zinc finger protein CONSTANS-LIKE 4 as the key candidate gene of stigma color in watermelon(Citrullus lanatus)

Stigma color is a critical agronomic trait in watermelon that plays an important role in pollination.However,there are few reports on the regulation of stigma color in watermelon.In this study,a genetic analysis of the F2 population derived from ZXG1553(P1,with orange stigma)and W1-17(P2,with yellow stigma)indicated that stigma color is a quantitative trait and the orange stigma is recessive compared with the yellow stigma.Bulk segregant analysis sequencing(BSA-seq)revealed a 3.75 Mb segment on chromosome 6 that is related to stigma color.Also,a major stable effective QTL Clqsc6.1(QTL stigma color)was detected in two years between cleaved amplified polymorphic sequencing(CAPS)markers Chr06_8338913 and Chr06_9344593 spanning a~1.01 Mb interval that harbors 51 annotated genes.Cla97C06G117020(annotated as zinc finger protein CONSTANS-LIKE 4)was identified as the best candidate gene for the stigma color trait through RNA-seq,quantitative real-time PCR(qRT-PCR),and gene structure alignment analysis among the natural watermelon panel.The expression level of Cla97C06G117020 in the orange stigma accession was lower than in the yellow stigma accessions with a significant difference.A nonsynonymous SNP site of the Cla97C06G117020 coding region that causes amino acid variation was related to the stigma color variation among nine watermelon accessions according to their re-sequencing data.Stigma color formation is often related to carotenoids,and we also found that the expression trend of ClCHYB(annotated asβ-carotene hydroxylase)in the carotenoid metabolic pathway was consistent with Cla97C06G117020,and it was expressed in low amounts in the orange stigma accession.These data indicated that Cla97C06G117020 and ClCHYB may interact to form the stigma color.This study provides a theoretical basis for gene fine mapping and mechanisms for the regulation of stigma color in watermelon.

Diagnostic value of methylated branched chain amino acid transaminase 1/IKAROS family zinc finger 1 for colorectal cancer

BACKGROUND The diagnostic value of combined methylated branched chain amino acid transaminase 1(BCAT1)/IKAROS family zinc finger 1(IKZF1)in plasma for colorectal cancer(CRC)has been explored since 2015.Recently,several related studies have published their results and showed its diagnostic efficacy.AIM To analyze the diagnostic value of methylated BCAT1/IKZF1 in plasma for screening and postoperative follow-up of CRC.METHODS The candidate studies were identified by searching the PubMed,Embase,Cochrane Library,CNKI,and Wanfang databases from May 31,2003 to June 1,2023.Sensitivity,specificity,and diagnostic accuracy were calculated by merging ratios or means.RESULTS Twelve eligible studies were included in the analysis,involving 6561 participants.The sensitivity of methylated BCAT1/IKZF1 in plasma for CRC diagnosis was 60%[95%confidence interval(CI)53-67]and specificity was 92%(95%CI:90-94).The positive and negative likelihood ratios were 8.0(95%CI:5.8-11.0)and 0.43(95%CI:0.36-0.52),respectively.Diagnostic odds ratio was 19(95%CI:11-30)and area under the curve was 0.88(95%CI:0.85-0.91).The sensitivity and specificity for CRC screening were 64%(95%CI:59-69)and 92%(95%CI:91-93),respectively.The sensitivity and specificity for recurrence detection during follow-up were 54%CONCLUSION The detection of methylated BCAT1/IKZF1 in plasma,as a non-invasive detection method of circulating tumor DNA,has potential CRC diagnosis,but the clinical application prospect needs to be further explored.

Therapeutic targeting of cellular prion protein: toward the development of dual mechanism anti-prion compounds

PrPSc,a misfolded,aggregation-prone isoform of the cellular prion protein(PrPC),is the infectious prion agent responsible for fatal neurodegenerative diseases of humans and other mammals.PrPSccan adopt different pathogenic conformations(prion strains),which can be resistant to potential drugs,or acquire drug resistance,posing challenges for the development of effective therapies.Since PrPCis the obligate precursor of any prion strain and serves as the mediator of prion neurotoxicity,it represents an attractive therapeutic target fo r prion diseases.In this minireview,we briefly outline the approaches to target PrPCand discuss our recent identification of Zn(Ⅱ)-Bn PyP,a PrPC-targeting porphyrin with an unprecedented bimodal mechanism of action.We argue that in-depth understanding of the molecular mechanism by which Zn(Ⅱ)-Bn PyP targets PrPCmay lead toward the development of a new class of dual mechanism anti-prion compounds.

Upregulated lncRNA PRNT promotes progression and oxaliplatin resistance of colorectal cancer cells by regulating HIPK2 transcription

BACKGROUND Colorectal cancer(CRC)is the third most common cancer and a significant cause of cancer-related mortality globally.Resistance to chemotherapy,especially during CRC treatment,leads to reduced effectiveness of drugs and poor patient outcomes.Long noncoding RNAs(lncRNAs)have been implicated in various pathophysiological processes of tumor cells,including chemotherapy resistance,yet the roles of many lncRNAs in CRC remain unclear.AIM To identify and analyze the lncRNAs involved in oxaliplatin resistance in CRC and to understand the underlying molecular mechanisms influencing this resistance.METHODS Gene Expression Omnibus datasets GSE42387 and GSE30011 were reanalyzed to identify lncRNAs and mRNAs associated with oxaliplatin resistance.Various bioinformatics tools were employed to elucidate molecular mechanisms.The expression levels of lncRNAs and mRNAs were assessed via quantitative reverse transcription-polymerase chain reaction.Functional assays,including MTT,wound healing,and Transwell,were conducted to investigate the functional implications of lncRNA alterations.Interactions between lncRNAs and trans-cription factors were examined using RIP and luciferase reporter assays,while Western blotting was used to confirm downstream pathways.Additionally,a xenograft mouse model was utilized to study the in vivo effects of lncRNAs on chemotherapy resistance.RESULTS LncRNA prion protein testis specific(PRNT)was found to be upregulated in oxaliplatin-resistant CRC cell lines and negatively correlated with homeodomain interacting protein kinase 2(HIPK2)expression.PRNT was demonstrated to sponge transcription factor zinc finger protein 184(ZNF184),which in turn could regulate HIPK2 expression.Altered expression of PRNT influenced CRC cell sensitivity to oxaliplatin,with overexpression leading to decreased sensitivity and decreased expression reducing resistance.Both RIP and luciferase reporter assays indicated that ZNF184 and HIPK2 are targets of PRNT.The PRNT/ZNF184/HIPK2 axis was implicated in promoting CRC progression and oxaliplatin resistance both in vitro and in vivo.CONCLUSION The study concludes that PRNT is upregulated in oxaliplatin-resistant CRC cells and modulates the expression of HIPK2 by sponging ZNF184.This regulatory mechanism enhances CRC progression and resistance to oxaliplatin,positioning PRNT as a promising therapeutic target for CRC patients undergoing oxaliplatin-based chemotherapy.

ZNF554 Inhibits Endometrial Cancer Progression via Regulating RBM5 and Inactivating WNT/β-Catenin Signaling Pathway

Objective:Uterine corpus endometrial carcinoma(UCEC),a kind of gynecologic malignancy,poses a significant risk to women’s health.The precise mechanism underlying the development of UCEC remains elusive.Zinc finger protein 554(ZNF554),a member of the Krüppel-associated box domain zinc finger protein superfamily,was reported to be dysregulated in various illnesses,including malignant tumors.This study aimed to examine the involvement of ZNF554 in the development of UCEC.Methods:The expression of ZNF554 in UCEC tissues and cell lines were examined by qRT-PCR and Western blot assay.Cells with stably overexpressed or knocked-down ZNF554 were established through lentivirus infection.CCK-8,wound healing,and Transwell invasion assays were employed to assess cell proliferation,migration,and invasion.Propidium iodide(PI)staining combined with fluorescence-activated cell sorting(FACS)flow cytometer was utilized to detect cell cycle distribution.qRT-PCR and Western blotting were conducted to examine relative mRNA and protein levels.Chromatin immunoprecipitation assay and luciferase reporter assay were used to explore the regulatory role of ZNF554 in RNA binding motif 5(RBM5).Results:The expression of ZNF554 was found to be reduced in both UCEC samples and cell lines.Decreased expression of ZNF554 was associated with higher tumor stage,decreased overall survival,and reduced disease-free survival in UCEC.ZNF554 overexpression suppressed cell proliferation,migration,and invasion,while also inducing cell cycle arrest.In contrast,a decrease in ZNF554 expression resulted in the opposite effect.Mechanistically,ZNF554 transcriptionally regulated RBM5,leading to the deactivation of the Wingless(WNT)/β-catenin signaling pathway.Moreover,the findings from rescue studies demonstrated that the inhibition of RBM5 negated the impact of ZNF554 overexpression onβ-catenin and p-glycogen synthase kinase-3β(p-GSK-3β).Similarly,the deliberate activation of RBM5 reduced the increase inβ-catenin and p-GSK-3βcaused by the suppression of ZNF554.In vitro experiments showed that ZNF554 overexpression-induced decreases in cell proliferation and migration were counteracted by RBM5 knockdown.Additionally,when RBM5 was overexpressed,it hindered the improvements in cell proliferation and migration caused by reducing the ZNF554 levels.Conclusion:ZNF554 functions as a tumor suppressor in UCEC.Furthermore,ZNF554 regulates UCEC progression through the RBM5/WNT/β-catenin signaling pathway.ZNF554 shows a promise as both a prognostic biomarker and a therapeutic target for UCEC.

Myeloid zinc finger 1(MZF1) is the most important transcriptional factor for porcine follistatin promoter

Follistatin （FS） is a secreted protein, which was originally isolated from porcine follicular fluid. Expression of follistatin is tightly regulated during porcine growth and development. To study the essential transcriptional regions of the porcine FS promoter, ten primer pairs were designed to amplify segments with different lengths of the FS promoter from -1 800 to ＋16 bp. The products were then inserted into the pGL3-basic vector to analyze the relative luciferase activity. The results showed that the most remarkable changes of promoter activity were observed between constructs （-302/＋16 bp）-FS and （-180/＋16 bp）-FS （P〈0.01）. Further research showed that the reconstructed reporter plasmid lacking myeloid zinc finger 1 （MZF1） binding sequence had significantly decreased luciferase activity （P〈0.05）. Furthermore, the FS protein expression was significantly increased in PK15 cells while the MZF1 was overexpressed, suggesting that the short sequence ＂TCCCCACC＂ （the recognition site of transcription factor MZF1） was the most important for FS transcription activation in the porcine.

Expression Pattern Analysis of Zinc Finger Protein Genes in Wheat(Triticum aestivum L.) Under Phosphorus Deprivation

Zinc finger protein（ZFP） genes comprise a large and diverse gene family, and are involved in biotic and abiotic stress responses in plants. In this study, a total of 126 ZFP genes classified into various types in wheat were characterized and subjected to expression pattern analysis under inorganic phosphate（Pi） deprivation. The wheat ZFP genes and their corresponding GenBank numbers were obtained from the information of a 4×44K wheat gene expression microarray chip. They were confirmed by sequence similarity analysis and named based on their homologs in Brachypodium distachyon or Oriza sativa. Expression analysis based on the microarray chip revealed that these ZFP genes are categorized into 11 classes according to their gene expression patterns in a 24-h of Pi deprivation regime. Among them, ten genes were differentially up-regulated, ten genes differentially downregulated, and two genes both differentially up- and down-regulated by Pi deprivation. The differentially up- or down-regulated genes exhibited significantly more or less transcripts at one, two, or all of the checking time points（1, 6, and 24 h） of Pi stress in comparison with those of normal growth, respectively. The both differentially up- and down-regulated genes exhibited contrasting expression patterns, of these, TaWRKY70;5 showed significantly up-regulated at 1 and 6 h and down-regulated at 24 h whereas TaAN1AN20-8;2 displayed significantly upregulated at 1 h and downregulated at 6 h under deprivation Pi condition. Real time PCR analysis confirmed the expression patterns of the differentially expressed genes obtained by the microarray chip. Our results indicate that numerous ZFP genes in wheat respond to Pi deprivation and have provided further insight into the molecular basis that plants respond to Pi deprivation mediated by the ZFP gene family.

Genome-wide analysis of the CCCH zinc finger family in longan:Characteristic identification and expression profiles in Dimocarpus longan Lour

CCCH(C3 H) Zinc finger(Znf) transcription factors(TFs), as a novel type of Znf gene, regulate the expression of genes by binding to their mRNAs and play important roles in plant growth and development and abiotic stress resistance.Longan(Dimocarpous longan) is a tropical/subtropical fruit tree of great economic importance in Southeast Asia.However, genomic information on C3 H and their functions in longan are still unknown. In this study, a comprehensive analysis of the longan C3 H(DlC3 H) gene family was carried out. A total of 49 DlC3 H genes in three clades were identified from the longan genome database. Characteristics of the genes were analyzed with respect to gene structure,motif composition, phylogenetic tree and potential functions. The analysis of alternative splicing(AS) events suggested that AS events in DlC3 H genes were related to the transformation from longan non-embryonic to embryonic cultures.Promoter analysis indicated that most of the DlC3 H genes included cis-acting elements associated with hormones and stresses responses. Quantitative real-time PCR(qRT-PCR) analysis indicated that 26 of the 49 DlC3 Hs, which possess methyl jasmonate(MeJA) and abscisic acid(ABA) responsive cis-acting elements, showed differential expression patterns under treatment with ABA, MeJA and their endogenous inhibitors, suggesting that DlC3 Hs might be involved in the ABA and MeJA signaling pathways. The expression profiles of 17 of the 49 DlC3 Hs in non-embryonic callus and three tissues of embryonic cultures showed that only five of the 17 DlC3 Hs had the same expression trends as the FPKM trends in transcriptome data;the expression levels of DlC3 H07/14/16/36/49 in embryogenic callus and DlC3 H04/38 in globular embryos were high, suggesting that they have different functions in embryonic development. Further, we verified that DlC3 H01/03/05/11/19/39 were regulated by sRNAs by a modified 5’ RLM-RACE method. This study provides the first systematic analysis of C3 H genes in longan, and found that C3 H genes may be involved in hormone and stress responses, and somatic embryogenesis. Our preliminary investigation may provide clues to further studies on the characteristics and functions of this family in longan.

Expression and analysis of zinc fi nger family gene in Lenzites gibbosa

Zinc finger transcription factors play significant roles in the growth and development of plant and animal,but their function remains obscure in fungi.Lenzites gibbosa mycelia were extracted and sequenced by transcriptome analysis after growing on sawdust at different times to support mycelial growth of L.gibbosa in a nutrient matrix.Data bases used for analysis were the Kyoto encyclopedia of genes and genomes(KEGG)annotation,the cluster of orthologous groups of proteins(COG)and gene ontology(GO)annotation.Zinc finger class genes related to the growth and development of L.gibbosa were screened.GO annotation and enrichment analysis of diff erentially expressed genes were carried out.A total of 114.55 Gb Clean Data were obtained from the L.gibbosa transcriptome.The average Clean Data in each sample was 6.16 Gb.The relative efficiency of reads between each sample and the reference genome was 88.5%to 91.4%.The COG analysis showed that most zinc finger protein genes were related to replication,recombination and repair function.GO enrichment analysis showed that the expressed genes involved in cellular process,cell part and binding.We identifi ed seventy-two expressed genes including seven up-regulated genes and sixty-five down-regulated genes by applying DESeq2 data analysis software.By comparing the significantly expressed genes with KEGG database,66 annotated sequences were obtained,and 35 primary metabolic pathways were annotated.Pathway enrichment analysis showed that differentially expressed genes were signifi cantly enriched in protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis pathways.Gene_11750 and gene_5266 are highly correlated with the growth and development of L.gibbosa and are closely related to protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis pathway.According to gene functional analysis,seven important differentially expressed genes related to the growth and development of L.gibbosa were identified.

PHOSPHORYLATION OF THE MYELOID ZINC FINGER PRO TEIN MZF－1 IS DIFFERENTIALLY REGULATED DURING MYELOPOIESIS

The myeloid zinc finger gene-1 (MZF-1) encodes a putative transcription factor whose expression has been implicated in myeloid differentiation. To study the role of the nMZF-1 in myploid differentiation,we characterized MZF-1 protein expr.ession,cellular localization,and phosphorylation in leukemia cell lines and leukemia cells.MZF1 protein expression was found only in myeloid cells. In proliferating HL-60 cells,MZF-1 was localized to the nucleus with some cytoplasmic distribution; however,upon retinoic acid (RA)induced granulocytic differentiation, MZF-1 became restricted to the nucleus.In32 PO4-la labelled HL-60 cell, MZF-1 was immunoprecipitated as a phosphoprotein doublet of 53￣54kDa. MZF-1 phosphorylation increased after acute stimulation of HL-60 with granulocytemacrophage colony stimulating factor (GM-CSF), interleukin-3(IL-3),phorbol ester,and serum.Chronic GM-CSf treatment of HL-60 cells potentiating granulocytic differentiation sustained the hyperphosphorylated state of MZF-1,whereas chronic treatment with TPA leading to monocytic-macrophage differentiation was accompanied by the disappearance of the 53 kDa MZF-1 phosphoprotein and the appearance of cross-reactive 69 and 105kDa phosphoprotein species. K562 human myeloblastic cells which are resistant to granulocytic differentiation express both the 53 kDa MZF-1 protein and the cross reactive 69 and 105 kDa proteins,but the 53 kDa MZF-1 protein is not detectable phosphorylated under any experimental conditions. Acute promyelocytic leukemic cells exhibited the 53kDa phosphoprotein,whereas monocytic leukemia cells expressed only the 69 and 105 kDa MZF-1 related phosphoproteins. The studies demonstrate that MZF-1 is a nuclear protein whose phosphorylation is associated with the granulocytic commitment of myeloid cells.

Molecular Cloning and Expression Analysis of a Cys2/His2 Type Zinc Finger Protein Gene in Upland Cotton

The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating

STUDIES ON THE SYNTHESIS AND DNA-BINDING ABILITY OF ZINC FINGER MOTIF

Transcription factor SPI is a protcin present in mammalian cells that binds to GC box promoter clements of Gene and selectively activates mRNA synthesis. The gene contains functional recognition sites. It contains three continuous zinc finger motifs, which are believed being mctalloprotein structures that interact with DNA. We synthesized the second zine finger fragment of SP1 (SP1-ZF2) and its mutant (SP1-ZF2 / HT. E20→H. R23→T), we also synthesized the Cys-Cys loop (ZF6) and the His-His loop (ZF5) of SPI and linked the twoloops together using a β-turn structure to obtain a finger mimic analogue (ZF-15) by stepwise solid-phase technique. Atomic absorption studies show that SP 1-ZF2 and SP1-ZF2 / HT bind zinc cquimolarly, but ZF-15 docs not bind Zn anyway. The CD experiments demonstrate a significant change in secondary structure in the prescnce or absence of Zn to SP1-ZF2 and SP1-ZF2/ HT, but there is no change about ZF-15. Gcl-retardation clectrophoresis assays indicate that SP1-ZF2 binds to DNA sequence specifically in the presence of Zn, but SP1-ZF2 / HT docs not bind as SP 1-ZF2 did. We observed that a single zine finger like SP1-ZF2 is able to bind DNA sequence specifically.

Long non-coding RNA-ATB induces trastuzumab resistance and aggravates the progression of gastric cancer by repressing miR- 200c via ZNF217 elevation

Background: Trastuzumab resistance accounts for chemotherapy failure in gastric cancer patients in clinicalpractice. The significance of long non-coding RNAs (lncRNAs) in the maintenance of drug resistance in gastriccancer has been already underlined. Method: This study aimed to identify the specific role of lncRNA-ATB in gastriccancer progression and trastuzumab resistance. The downstream miRs of lncRNA-ATB and target genes of miRs werepredicted by bioinformatics analysis and verified using dual luciferase reporter assay. Loss- and gain-function assayswere performed to explore the roles of lncRNA-ATB, miR-200c, and zinc-finger protein 217 (ZNF217) in the cellfunctions and trastuzumab resistance of a trastuzumab-resistant gastric cancer cell line (NCI-N87-TR). Result:LncRNA-ATB was upregulated, while miR-200c was downregulated. Depletion of lncRNA-ATB or miR-200celevation led to a decrease in malignant properties of NCI-N87-TR cells. LncRNA-ATB could negatively target miR-200c, which in turn inversely targeted and reduced the expression of ZNF217. Silencing of ZNF217 could inhibit cellviability and migration. Conclusion: lncRNA-ATB promoted the progression and trastuzumab resistance of gastriccancer by repressing miR-200c via ZNF217 upregulation.

Clinicopathological Significance of Increased ZIC1 Expression in Human Endometrial Cancer

Summary： Zinc finger of the cerebellum （ZIC 1）, one of ZIC family genes, has been shown to play im- portant roles in many cancers such as gastric cancer and breast cancer. However, there is little known about the expression and significance of ZIC1 in endometrial cancer. The aim of this study was to de- termine the expression pattern and clinicopathological significance of ZIC1 in endometrial cancer. The rnRNA and protein expression of ZIC1 in endometrial cancer tissues was detected using the reverse- transcription polymerase chain reaction and Western blotting, respectively. Immunostaining of ZIC1 in 99 endometrial cancer samples was examined and its associations with clinicopathological parameters were analyzed. Hec-l-B cells were transfected with Z1CI-shRNA or sc-shRNA, and cell proliferation was assayed. Hec-l-B cells stably transfected with ZICI-shRNA or sc-shRNA were subcutaneously in- oculated into nude mice, and the tumor weight was measured. A significantly increased expression of ZIC1 mRNA and protein was observed in endometrial cancer tissues compared to that in normal endo- metrial tissues （P〈0.05）. Immunohistochemical analysis showed that strong cytoplasmic immunostain- ing of ZIC1 was observed in almost all endometrial cancer samples （90/99） while light and moderate immunostaining of ZIC 1 was only detected in 17 of 30 （56.7%） normal tissues. Moreover, up-regulation of ZIC1 was significantly correlated with age, disease stage, TNM stage and FIGO stage （P〈0.05）. The down-regulated expression of ZIC1 contributed to the inhibition of cell proliferation, and inhibited the growth of tumor. It was concluded that ZIC1 is over-expressed in endometrial cancer tissue but not in normal tissue, and positively correlated to the malignant biological behavior of endometrial carcino- genesis.

Application of Artificially Induced Double-strand Breaks (DSB) and Triplex-forming Oligonucleotides (TFO) in the Improvement of Gene Targeting Efficiency

Gene targeting technology is an important means to investigate gene functions, but its efficiency of gene targeting is very low, especially for somatic cell targeting. Artificially induced double-strand breaks （DSB） and triplex forming oligonucleotide （TFO） are currently developed methods to improve the targeting efficiency. This paper summarized the basic principles, design ideas and application in gene targeting efficiency improvement of these two methods, analyzed and com- pared their characteristics, and finally proposed prospects for their future development.

鼻咽癌组织中WNT8B与ZNF191蛋白表达与临床病理及预后的相关性分析

目的 探讨鼻咽癌组织中Wnt家族成员8B(Wnt family member 8B,WNT8B)与锌指转录因子191(zinc finger transcription factor 191,ZNF191)蛋白表达与临床病理及预后的相关性。方法 收集2017年1月~2019年1月东阳市人民医院保存的126例鼻咽癌组织石蜡标本及其临床病理资料(病例组),另选取慢性鼻咽炎组织样本40例(对照组)。采用免疫组织化学法检测2组样本组织中WNT8B和ZNF191蛋白表达情况,并对比两蛋白阳性率及共表达阳性率,分析病例组患者鼻咽癌组织中WNT8B和ZNF191蛋白表达与临床病理因素关系,采用Kaplan-Meier法和Cox回归分析法分析鼻咽癌组织中WNT8B和ZNF191表达与预后的关系。结果 病例组鼻咽癌组织中WNT8B和ZNF191阳性率及二者共表达阳性率均高于对照组鼻咽黏膜组织(P<0.05)。Spearson相关性分析结果显示,鼻咽癌组织中WNT8B与ZNF191表达呈正相关(P<0.05)。T3~T4级、Ⅲ~Ⅳ期、有淋巴结转移的鼻咽癌组织中WNT8B和ZNF191阳性率及二者共表达阳性率分别高于T1~T2级、Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05)。WNT8B阳性与阴性患者3年生存率分别为57.89%、83.87%,ZNF191阳性与阴性患者分别为56.79%、77.78%,共表达阳性患者与非共表达患者分别为50.75%、79.66%;经Kaplan-Meier分析结果显示,WNT8B阳性患者与阴性、ZNF191阳性与阴性、共表达阳性与非共表达患者累积存活曲线比较,差异均有统计学意义(P<0.05)。Cox回归分析显示,浸润深度[HR(95%CI)=1.980(1.023~3.955)]、TNM分期[HR(95%CI)=3.207(1.936~11.751)]、WNT8B阳性表达[HR(95%CI)=4.506(2.068~13.774)]、ZNF191阳性表达[HR(95%CI)=3.114(2.061~10.485)]、WNT8B+ZNF191共表达[HR(95%CI)=4.691(2.144~14.902)]是影响总生存期的独立危险因素(P<0.05)。结论 鼻咽癌患者鼻咽癌组织中WNT8B和ZNF191阳性表达率高,WNT8B和ZNF191表达与鼻咽癌患者浸润深度、TNM分期、淋巴结转移及预后生存相关。

In Vivo Modeling of Zebrafish Zinc Finger,MIZ-Type Containing 1 Expression and Its Effect on Pigmentation

Objective:The zinc finger,MIZ-type containing 1(ZMIZ1)gene has been identified as a possible susceptibility gene associated with vitiligo,therefore we conducted this study to investigate the role ofZMIZ1 in pigmentation.Methods:We generate a zebrafish loss-of-function model using morpholino oligonucleotides(MOs),and two orthologs of humanZMIZ1 have been annotated(ZMIZ1a andZMIZ1b).The expression profiles of ZMIZ1a and ZMIZ1b and their effects on the pigmentation in zebrafish were evaluated by using whole-mount in situ hybridization and melanin quantification.Statistical analysis was performed using the unpaired Studentt-test or one-way analysis.Results:Investigation of the temporal and spatial expressions of these two transcripts suggested that the expressions ofZMIZ1a andZMIZ1b in the brain start to emerge in a ubiquitous fashion from 2 days post-fertilization onwards.After the successful design and validation of MOs,we observed thatZMIZ1a andZMIZ1b MOs caused embryonic developmental delays and malformations in zebrafish.Further analysis of the melanin content in the morphants revealed thatZMIZ1a significantly(49.1%for 0.667 mmol/L inZMIZI1a group,P=0.03)reduced the melanin content in a dose-dependent manner,but only the highest concentration of injectedZMIZ1b MOs significantly(50%for 0.667 mmol/L inZMIZ1b group,P=0.02)reduced the melanin content.A tyrosinase inhibition assay indicated no significant difference between the morphants and wild-type zebrafish.Conclusion:This study successfully modeled a susceptibility gene identified by genome-wide association studies in a zebrafish loss-of-function model and provides insights into the biological mechanism of pigmentation.

Mitochondrial diseases and mtDNA editing

Mitochondrial diseases are a heterogeneous group of inherited disorders character-ized by mitochondrial dysfunction,and these diseases are often severe or even fatal.Mito-chondrial diseases are often caused by mitochondrial DNA mutations.Currently,there is no curative treatment for patients with pathogenic mitochondrial DNA mutations.With the rapid development of traditional gene editing technologies,such as zinc finger nucleases and tran-scription activator-like effector nucleases methods,there has been a search for a mitochon-drial gene editing technology that can edit mutated mitochondrial DNA;however,there are still some problems hindering the application of these methods.The discovery of the DddA-derived cytosine base editor has provided hope for mitochondrial gene editing.In this paper,we will review the progress in the research on several mitochondrial gene editing technologies with the hope that this review will be useful for further research on mitochondrial gene editing technologies to optimize the treatment of mitochondrial diseases in the future.

ZiF-Predict:A Web Tool for Predicting DNA-Binding Specificity in C2H2 Zinc Finger Proteins

Engineering zinc finger protein motifs for specific DNA targets in genomes is critical in the field of genome engineering. We have developed a computational method for predicting recognition helices for C2H2 zinc fingers that bind to specific target DNA sites. This prediction is based on artificial neural network using an exhaustive dataset of zinc finger proteins and their target DNA triplets. Users can select the option for two or three zinc fingers to be predicted either in a modular or synergistic fashion for the input DNA sequence. This method would be valuable for researchers interested in designing specific zinc finger transcription factors and zinc finger nucleases for several biological and biomedical applications. The web tool ZiF-Predict is available online at http：//web.iitd.ac.in/-sundar/zifpredict/.

The C_2H_2 -type Zinc Finger Protein ZFP182 is Involved in Abscisic Acid-Induced Antioxidant Defense in Rice

C2H2-type zinc finger proteins （ZFPs） are thought to play important roles in modulating the responses of plants to drought, salinity and oxidative stress. However, direct evidence is lacking for the involvement of these ZFPs in abscisic acid （ABA）-induced antioxidant defense in plants. In this study, the role of the rice （Oryza sativa L. sub.japonica cv. Nipponbare） C2H2-type ZFP ZFP182 in ABA-induced antioxidant defense and the relationship between ZFP182 and two rice MAPKs, OsMPK1 and OsMPK5 in ABA signaling were investigated. ABA treatment induced the increases in the expression of ZFP182, OsMPK1 and OsMPK5, and the activities of superoxide dismutase （SOD） and ascorbate peroxidase （APX） in rice leaves. The transient gene expression analysis and the transient RNA interference （RNAi） analysis in protoplasts showed that ZFP182, OsMPK1 and OsMPK5 are involved in ABA-induced up-regulation in the activities of SOD and APX. Besides, OsMPK1 and OsMPK5 were shown to be required for the up-regulation in the expression of ZFP182 in ABA signaling, but ZFP182 did not mediate the ABA-induced up-regulation in the expression of OsMPK1 and OsMPKS. These results indicate that ZFP182 is required for ABA-induced antioxidant defense and the expression of ZFP182 is regulated by rice MAPKs in ABA signaling.