Publications by authors named "Changying Niu"

21 Publications

  • Page 1 of 1

Mutation of P-element somatic inhibitor induces male sterility in the diamondback moth, Plutella xylostella.

Pest Manag Sci 2021 Apr 11. Epub 2021 Apr 11.

CAS Key Laboratory of Insect Developmental and Evolutionary Biology, CAS Center for Excellence in Molecular Plant Sciences/Institute of Plant Physiology and Ecology, Shanghai, China.

Background: Genetic manipulation of sex determination pathways in insects provides the basis for a broad range of strategies to benefit agricultural security and human health. The P-element somatic inhibitor (PSI) protein, an exon splicing silencer that promotes male-specific splicing of dsx, plays a critical role in male sexual differentiation and development. The functions of PSI have been characterized in the lepidopteran model species Bombyx mori. However, the molecular mechanism and functions of PSI in Plutella xylostella, a worldwide agricultural pest and taxonomically basal species, are still unknown.

Results: Here we identified PxPSI transcripts and analyzed their spatiotemporal expression pattern in P. xylostella. Multiple sequence alignment revealed that PxPSI contains four KH domains and is highly conserved in lepidopterans. We used the CRISPR-Cas9 system to generate mutations of the PxPSI genomic locus. Disruptions of PxPSI caused male-specific defects in internal and external genitals. In addition, we detected female-specific Pxdsx transcripts in PxPSI male mutants. Mutations also caused changes in expression of several sex-biased genes and induced male sterility.

Conclusion: Our study demonstrates that PxPSI plays a key role in male sex determination in P. xylostella and suggests a potential molecular target for genetic-based pest management in lepidopteran pests.
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http://dx.doi.org/10.1002/ps.6413DOI Listing
April 2021

Identification and functional characterization of odorant-binding proteins 69a and 76a of .

Heliyon 2021 Mar 10;7(3):e06427. Epub 2021 Mar 10.

Beijing Key Laboratory of Environment Friendly Management on Fruit Diseases and Pests in North China, Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing, China.

The fruit fly is a fruit crop pest that causes a severe economic threat to soft summer fruit worldwide. The male sex pheromone, cis-vaccenyl acetate (cVA) has multiple functions in intra-species communication in , which is required in male to suppress male-male courtship. males do not produce cVA; however, the odorant receptor for cVA (Or67d) is still functional. The lack of cVA in casts the question of whether this pheromone might have been replaced by another compound similar to cVA that disrupts mating in . In order to address this question, we cloned two adult antenna-specific odorant-binding proteins (OBPs) DsOBP69a and DsOBP76a and aligned with their orthologues. Moreover, we examined the binding properties of the newly identified recombinant proteins against 26 potential ligands including cVA, using the fluorescence-based ligand binding assay. The alignment showed that DsOBP69a and DsOBP76a, have six conserved cysteines and belong to the classic OBP family. Furthermore, our results revealed that cVA did not bind to DsOBP69a or DsOBP76a proteins. Interestingly, the floral odorant β-ionone and the bitter substance berberine chloride and coumarin displayed high binding ability. It is also worth noting that DsOBP69a and DsOBP76a have different affinities to (Z)-7-Tricosene that may reflect different functional roles. These findings suggest that and are potentially involved in olfaction and gustation of .
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http://dx.doi.org/10.1016/j.heliyon.2021.e06427DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7970147PMC
March 2021

Detection and characterization of bacterial endosymbionts in Southeast Asian tephritid fruit fly populations.

BMC Microbiol 2019 12 24;19(Suppl 1):290. Epub 2019 Dec 24.

Department of Environmental Engineering, University of Patras, 2 Seferi St., 30100, Agrinio, Greece.

Background: Various endosymbiotic bacteria, including Wolbachia of the Alphaproteobacteria, infect a wide range of insects and are capable of inducing reproductive abnormalities to their hosts such as cytoplasmic incompatibility (CI), parthenogenesis, feminization and male-killing. These extended phenotypes can be potentially exploited in enhancing environmentally friendly methods, such as the sterile insect technique (SIT), for controlling natural populations of agricultural pests. The goal of the present study is to investigate the presence of Wolbachia, Spiroplasma, Arsenophonus and Cardinium among Bactrocera, Dacus and Zeugodacus flies of Southeast Asian populations, and to genotype any detected Wolbachia strains.

Results: A specific 16S rRNA PCR assay was used to investigate the presence of reproductive parasites in natural populations of nine different tephritid species originating from three Asian countries, Bangladesh, China and India. Wolbachia infections were identified in Bactrocera dorsalis, B. correcta, B. scutellaris and B. zonata, with 12.2-42.9% occurrence, Entomoplasmatales in B. dorsalis, B. correcta, B. scutellaris, B. zonata, Zeugodacus cucurbitae and Z. tau (0.8-14.3%) and Cardinium in B. dorsalis and Z. tau (0.9-5.8%), while none of the species tested, harbored infections with Arsenophonus. Infected populations showed a medium (between 10 and 90%) or low (< 10%) prevalence, ranging from 3 to 80% for Wolbachia, 2 to 33% for Entomoplasmatales and 5 to 45% for Cardinium. Wolbachia and Entomoplasmatales infections were found both in tropical and subtropical populations, the former mostly in India and the latter in various regions of India and Bangladesh. Cardinium infections were identified in both countries but only in subtropical populations. Phylogenetic analysis revealed the presence of Wolbachia with some strains belonging either to supergroup B or supergroup A. Sequence analysis revealed deletions of variable length and nucleotide variation in three Wolbachia genes. Spiroplasma strains were characterized as citri-chrysopicola-mirum and ixodetis strains while the remaining Entomoplasmatales to the Mycoides-Entomoplasmataceae clade. Cardinium strains were characterized as group A, similar to strains infecting Encarsia pergandiella.

Conclusions: Our results indicated that in the Southeast natural populations examined, supergroup A Wolbachia strain infections were the most common, followed by Entomoplasmatales and Cardinium. In terms of diversity, most strains of each bacterial genus detected clustered in a common group. Interestingly, the deletions detected in three Wolbachia genes were either new or similar to those of previously identified pseudogenes that were integrated in the host genome indicating putative horizontal gene transfer events in B. dorsalis, B. correcta and B. zonata.
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http://dx.doi.org/10.1186/s12866-019-1653-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7050614PMC
December 2019

Mutation of the seminal protease gene, serine protease 2, results in male sterility in diverse lepidopterans.

Insect Biochem Mol Biol 2020 01 18;116:103243. Epub 2019 Sep 18.

Key Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, 200032, Shanghai, China. Electronic address:

Sterile insect technology (SIT) is an environmentally friendly method for pest control. As part of our efforts to develop a strategy that results in engineered male-sterile strains with minimum effects on viability and mating competition, we used CRISPR/Cas9 technology to disrupt Ser2, which encodes a seminal fluid protein, in the model lepidopteran insect, Bombyx mori, and an important agricultural pest, Plutella xylostella. Disruption of Ser2 resulted in dominant heritable male sterility. Wild-type females mated with Ser2-deficient males laid eggs normally, but the eggs did not hatch. We detected no differences in other reproductive behaviors in the mutant males. These results support the conclusion that Ser2 gene is necessary for male reproductive success in diverse lepidopterans. Targeting Ser2 gene has the potential to form the basis for a new strategy for pest control.
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http://dx.doi.org/10.1016/j.ibmb.2019.103243DOI Listing
January 2020

Mutation of Bdpaired induces embryo lethality in the oriental fruit fly, Bactrocera dorsalis.

Pest Manag Sci 2020 Mar 4;76(3):944-951. Epub 2019 Oct 4.

Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control, College of Plant Science & Technology, Huazhong Agricultural University, Wuhan, China.

Background: Pair-rule genes were identified and named for their role in segmentation in animal embryos. Paired, a homolog of mammalian PAX3, acts as one of several pair-rule genes and is key in defining the boundaries of future parasegments and segments during insect embryogenesis.

Results: We cloned the paired gene from the oriental fruit fly, Bactrocera dorsalis, and then applied CRISPR/Cas9-mediated genome editing to investigate its physiological function in the embryonic stage of this pest. We identified one transcript for a paired homolog in B. dorsalis, which encodes a protein containing a Paired Box domain and a Homeobox domain. Phylogenetic analysis confirmed that the paired gene is highly conserved and the gene was highly expressed at the 12-14 h-old embryonic stage. Knock-out of Bdpaired led to lack of segment boundaries, cuticular deficiency, and embryonic lethality. Sequence analysis of the CRISPR/Cas9 mutants exhibited different insertion and deletions in the Bdpaired locus. In addition, the relative expression of Wingless (Wg) and Abdominal A (Abd-A) genes were significantly down-regulated in the Bdpaired mutant embryos.

Conclusion: These results indicate that Bdpaired gene is critical for the embryonic development of B. dorsalis, and could be a novel molecular target for genetic-based pest management practices to combat this serious invasive pest. © 2019 Society of Chemical Industry.
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http://dx.doi.org/10.1002/ps.5602DOI Listing
March 2020

Fermented crop straws by Trichoderma viride and Saccharomyces cerevisiae enhanced the bioconversion rate of Musca domestica (Diptera: Muscidae).

Environ Sci Pollut Res Int 2019 Oct 9;26(28):29388-29396. Epub 2019 Aug 9.

Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control, College of Plant Science & Technology, Huazhong Agricultural University, Wuhan, 430070, China.

Crop straw is an abundant renewable resource whose usage is limited due to its high cellulose, hemicellulose, and lignin contents. Here, Trichoderma viride, Saccharomyces cerevisiae, and Musca domestica were used to transform crop straws, and we investigated their impact on housefly rearing performance and optimized their utilization. The weights of cellulose, hemicellulose, and lignin in fermented crop straw diets significantly decreased after bioconversion by M. domestica larvae. The highest bioconversion rate was recorded in corn straw diet (16.19%), followed by wheat straw diet (10.31%) and wheat bran diet (8.97%). Similarly, high larval weight (yield) and pupation rate and fecundity and fertility rate were recorded in fermented crop straw diets composed of corn straw and wheat bran in 1:1 proportions. These results indicated that fermenting crop straw with T. viride and S. cerevisiae represented an efficient strategy that enhanced crop straw bioconversion and improved the rearing capacity of the housefly larvae. The resulting larvae could further be used as proteinaceous feed in poultry and aquaculture industries. Graphical abstract.
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http://dx.doi.org/10.1007/s11356-019-06101-1DOI Listing
October 2019

Mutation of doublesex induces sex-specific sterility of the diamondback moth Plutella xylostella.

Insect Biochem Mol Biol 2019 09 3;112:103180. Epub 2019 Jul 3.

CAS Key Laboratory of Insect Developmental and Evolutionary Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai, 200032, China. Electronic address:

DOUBLESEX (DSX): the downstream gene in the insect sex determination pathway, plays a critical role in sexual differentiation and development. The functions of dsx have been characterized in several model insect species. However, the molecular mechanism and functions of sex determination of dsx in Plutella xylostella, an agricultural pest, are still unknown. In present study, we identified a male-specific and three female-specific Pxdsx transcripts in P. xylostella. Phylogenetic analyses and multiple sequence alignment revealed that Pxdsx is highly conserved in lepidopterans. The CRISPR/Cas9 technology was used to induce mutations in the male-specific isoform, the female-specific isoform, and common regions of Pxdsx. Disruptions of Pxdsx sex-specific isoforms caused sex-specific defects in external genitals and partial sexual reversal. In addition, we found that female specific transcripts were detected in Pxdsx male mutants and male-specific transcripts were detected in Pxdsx female mutants. Mutations also caused changes in expression of several sex-biased genes and induced sex-specific sterility. This study demonstrates that Pxdsx plays a key role in sex determination of P. xylostella and suggests novel genetic control approaches for the management of P. xylostella.
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http://dx.doi.org/10.1016/j.ibmb.2019.103180DOI Listing
September 2019

Development of an easy and cost-effective method for non-invasive genotyping of insects.

PLoS One 2019 3;14(6):e0216998. Epub 2019 Jun 3.

Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science & Technology, Huazhong Agricultural University, Wuhan, Hubei, China.

Non-invasive genotyping methods provide valuable information on insect populations. However, poor DNA amplification and time-consuming sampling procedures limit these methods, especially for small insects. An efficient and convenient method was developed for non-invasive, non-lethal genotyping of a large insect, Mythimna separata, and a small insect, Drosophila melanogaster, by amplification of endogenous and exogenous, nuclear and mitochondrial genes from insect frass, exuviae, and food waste. For M. separata, the chitin synthesis gene MsCHSB and the COI gene were successfully detected by PCR from exuviae DNA. However, a COI fragment could not be detected directly by PCR from frass, probably due to DNA degradation. To improve the detection rate, DNA from frass was first amplified by Multiple Displacement Amplification with phi29 DNA polymerase, after which the COI fragment was detected from all samples by PCR. For D. melanogaster, second instar larvae were reared individually for three days and then DNA was extracted from food waste of each individual. The endogenous fragment serendipity α (sryα), exogenous transgene ΦC31 integrase, and the kl-5 gene, a Y-chromosome-located male-specific marker gene were successfully detected from most samples. We developed a simple, non-invasive, non-lethal method to determine gender and identify transgenic individuals early in the larval stage. This universal method is applicable to most insects and has potential application in genetic and ecological studies of insects and other arthropods.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0216998PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6546216PMC
February 2020

Rh6 gene modulates the visual mechanism of host utilization in fruit fly Bactrocera minax.

Pest Manag Sci 2019 Jun 28;75(6):1621-1629. Epub 2018 Dec 28.

Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control, College of Plant Science & Technology, Huazhong Agricultural University, Wuhan, China.

Background: Vision plays a critical role in host location and oviposition behavior for herbivorous insects. However, the molecular mechanisms underlying visual regulation in host recognition and oviposition site selection in insects remains unknown. The aim of this study was to explore the key visual genes that are linked to the host plant location of the fruit fly, Bactrocera minax.

Results: Using a host specialist fruit fly, B. minax, which lays eggs only into immature green citrus fruit, we undertook behavioral, transcriptomic, and RNAi research to identify the molecular basis for host fruit color recognition. In laboratory and field assays we found that adults prefer green over other colors, and this preference is significantly increased in sexually mature over immature flies. Furthermore, we identified that the Rh6 gene, responsible for green spectral sensitivity, has elevated expression in mature flies over immature flies. RNAi suppression of Rh6 eliminated the preference for green, resulting in a significant decrease in the number of eggs laid by B. minax in green unripe citrus.

Conclusion: These results show that the Rh6 gene modulates the visual mechanism of host utilization in B. minax, providing a genetic basis for visual host location in a non-model insect herbivore. © 2018 Society of Chemical Industry.
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http://dx.doi.org/10.1002/ps.5278DOI Listing
June 2019

[EFFECTIVENESS OF V-Y ADVANCED RETROAURICULAR FLAP FOR REPAIRING MILD AND MODERATE EARLOBE DEFECTS].

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2016 Nov;30(11):1370-1372

Institute of Reconstructive Plastic Surgery, Weifang Medical University, Weifang Shandong, 261042, P.R.China.

Objective: To investigate the effectiveness of the V-Y advanced retroauricular flap for repairing mild and moderate earlobe defect.

Methods: Between September 2014 and July 2015, V-Y advanced retroauricular flap was used to repair earlobe defect in 6 patients. There were 1 male and 5 females, aged 18 to 30 years (mean, 23 years). The left earlobe was involved in 2 cases and the right earlobe in 4 cases, including 2 cases of congenital earlobe defect and 4 cases of secondary earlobe defect; 1 patient had congenital deformity of upper auricle. According to self-made criteria for earlobe defect, 5 cases were rated as mild defect and 1 case as moderate defect.

Results: All incisions healed at the first stage, and the flaps survived smoothly. The patients were followed up 3 to 12 months, with an average of 9 months. The reconstructed earlobes had natural size and shape, and smooth curve; the texture and color were close to the adjacent skin. The effectiveness was satisfactory.

Conclusions: The V-Y advanced retroauricular flap for repairing mild and moderate earlobe defect can achieve natural earlobe and aesthetic plastic effectiveness, so it is a safe and ideal earlobe reconstruction method.
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http://dx.doi.org/10.7507/1002-1892.20160280DOI Listing
November 2016

Pupal diapause termination in Bactrocera minax: an insight on 20-hydroxyecdysone induced phenotypic and genotypic expressions.

Sci Rep 2016 06 8;6:27440. Epub 2016 Jun 8.

College of Plant Science &Technology, Huazhong Agricultural University, Wuhan 430070, China.

The Chinese citrus fruit fly, Bactrocera minax, is an economically important pest of citrus. It exhibits pupal diapause from November to May to combat harsh environmental conditions. Such a long pupal diapause is a barrier for laboratory rearing and development of control strategies against this pest. In the present study, 20-hydroxyecdysone (20E) was used to break pupal diapause of B. minax by topical application. After diapause termination by 20E treated, the pupal ontogenetic processes were observed along the temporal trajectory. The pupal response time to 20E was estimated by detecting the relative expression of 20E responsive genes at different times after 20E-treatment. Results revealed that 20E could effectively terminate the pupal diapause in a dose-dependent manner and significantly shorten the time for 50% adult emergence (Et50). 20E response genes, including ecr, broad and foxo, were up-regulated within 72h, indicating these genes are involved in pupal metamorphosis and diapause termination processes. Morphological changes showed the pupal metamorphosis began ~7 days after 20E-treatment at 22 °C. This study does not only pave the way for artificial rearing in the laboratory through manipulating of pupal diapause termination, but also deepens our understanding of the underlying pupal diapause termination mechanism of B. minax.
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http://dx.doi.org/10.1038/srep27440DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4897610PMC
June 2016

[The experimental study of the effect of ASCs on the skin expansion rate in rabbit].

Zhonghua Zheng Xing Wai Ke Za Zhi 2016 Mar;32(2):136-41

Objective: To explore the effect of adipose-derived stem cells (ASCs) on the skin expansion rate in rabbit.

Methods: The rabbit ASCs were isolated from fat tissue and cultured in vitro. The ADSCs were identified by cell immunofluorescence and marked by Edu staining.20 new Zealand rabbits were randomly divided into experimental(n =10) and control group(n =10).An area of 1.5 cm ×1.5 cm on the one side back of each rabbit was tattooed and one 30 ml round expander was implanted subcutaneously. ASCs suspension (1 ml) was injected subcutaneously in the experimental group, while serum free DMEM medium(1 ml) in control group. The expansion was proceeded regularly under constant pressure for 4 weeks.The expanded tattooed square area was measured on the 7th,14th,28th day and analyzed statistically. The expanded skin was harvested for histological study. Immunohistochemical staining was used to detect the expression of vascular endothelial cell marker CD31,and the microvessel density determination. The expression of epidermal growth factor (EGF) and vascular endothelial growth factor(VEGF)was detected by ELISA for skin tissue specificity. Western Blot was used for detection of CK19 in the epidermal cells.

Results: The expanded skin thickness and expansion rate in experimental group were significant higher than those in control group (P < 0.05). Compared with control group, the expression of CK19,CD31 and EGF, VEGF, as well as the microvessel density were all markedly increased in experimental group(P <0.05).

Conclusions: ASCs can increase the expansion rate of skin tissue by promotion of angiogenesis and tissue regeneration.
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March 2016

[The feasibility of adenoviral co-transduction of BMP2 and BMP7 for the expression of recombinant human BMP2/7 heterodimer in rat bone marrow mesenchymal stem cells].

Zhonghua Zheng Xing Wai Ke Za Zhi 2016 Mar;32(2):125-29

Objective: To investigate the feasibility of rat bone marrow mesenchymal stem cells (BMSCs) as the target cell of adenovirus-mediated co-transduction of BMP2 and BMP7 genes and then facilitate the expression of recombinant BMP2/7 heterodimer protein.

Methods: 3 adult male Fischer 344 rats of about 10 weeks of age were used for harvest and in vitro culture of rat BMSCs. Recombinant adenovirus vector carrying BMP2 or BMP7 target genes were constructed with AdMax vector system, and production of high-titer adenoviruses were packaged with HEK293T cells and then concentrated with CSCl2 density-gradient ultra-centrifugation. Rat BMSCs from passage 3 were seeded in 6-well plates at the concentration of 10 000 cells/cm2.After overnight pre-culture, BMSCs were allowed to culture in 200 μl serum-free alpha MEM containing both Ad-BMP2 and Ad-BMP7 adenovirus (100 MOI of each virus). After 7 days in vitro culture, conditioned cell culture supernatants were collected and followed by immunoprecipitation through immune protein G columns pre-loaded with mouse anti-human BMP7 antibody. The resulted protein immune-precipitates were used to assay the expression of BMP2/7 heterodimers via Western Blot and ELISA assay. As a negative control, Rat BMSCs were also genetically transduced with Ad-GFP virus at a concentration of 200 MOI.

Results: Our data demonstrated that recombinant adenoviruses carrying BMP2 or BMP7 target gene was successfully reconstructed, packaged, and confirmed via Western Blot assay, which as respected, presented as an unique band at 55 000 size for BMP2 or 49 000 size for BMP7.Adenovirus Ad-GFP was used to verify the integrity of recombinant virus and its transfection efficiency in rat BMSCs, which showed well cell attachment to culture plate and had no cytotoxicity. Green fluorescent protein in BMSCs was also noted eminently under fluorescent microscope. Combined transduction with AdBMP2 plus Ad-BMP7 resulted in the formation of BMP2/7 heterodimers from rat BMSCs. Analysis of conditioned medium via Western Blot assay showed a single protein band of about 47 000 size, just as expected. Quantitative ELISA analysis presented a prominent expression of about (4.33 ± 0.42) ng/ml for recombinant BMP2/7 heterodimers. A paired t test showed significant difference (P < 0.05),when compare to control groups of (0.08 ± 0.02) ng/ml.

Conclusions: As an ideal cell source for tissue engineering, rat BMSCs can be genetically modified with Ad-BMP2 plus Ad-BMP7 mediated co-transduction strategy, and efficiently produce recombinant human BMP2/7 heterodimers in vitro, which should facilitate further studies on the beneficial effect of BMP2/7 heterodimers to ex vivo osteogenesis of BMSCs
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March 2016

[EFFECTIVENESS OF MODIFIED Park METHOD OF BLEPHAROPLASTY FOR CORRECTION OF MILD BLEPHAROPTOSIS].

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2015 Sep;29(9):1133-6

Objective: To explore the effectiveness of the modified Park method of blepharoplasty for correction of mild blepharoptosis.

Methods: Between October 2012 and January 2015, a new modified Park method of blepharoplasty was performed on 23 patients with foldless eyelid combined mild blepharoptosis. There were 14 males and 9 females, aged 16 to 35 years (mean, 25 years). Unilateral eyelid was involved in 16 cases, bilateral eyelids in 7 cases. The upper eyelid was located at the edge of the pupil, and the drop was 1-2 mm (mean, 1.5 mm).

Results: All incisions healed at the first stage; no obvious blood stasis and swelling occurred. The patients were followed up 4 to 26 months, with an average of 15 months. The double eyelid fold was natural and smooth, and ptosis was completely corrected; the eyelid shape and position were symmetry when in situ fixation and movement. According to "double eyelid operation effect evaluation standard discussion" method by Chinese Medical Cosmetology Association, the results were excellent in all patients.

Conclusion: The modified Park method of blepharoplasty can achieve blepharoplasty and correcting blepharoptosis at the same time for correction of foldless eyelid combined mild blepharoptosis during operation without separated and amputated levator aponeurosis, with small surgical trauma, good controllability, and maneuverability in correction amplitude.
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September 2015

Transgenic Cotton Plants Expressing Double-stranded RNAs Target HMG-CoA Reductase (HMGR) Gene Inhibits the Growth, Development and Survival of Cotton Bollworms.

Int J Biol Sci 2015 15;11(11):1296-305. Epub 2015 Sep 15.

College of Plant Science and Technology, National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei 430070, P.R. China.

RNA interference (RNAi) has been developed as a powerful technique in the research of functional genomics as well as plant pest control. In this report, double-stranded RNAs (dsRNA) targeting 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene, which catalyze a rate-limiting enzymatic reaction in the mevalonate pathway of juvenile hormone (JH) synthesis in cotton bollworm, was expressed in cotton plants via Agrobacterium tumefaciens-mediated transformation. PCR and Sothern analysis revealed the integration of HMGR gene into cotton genome. RT-PCR and qRT-PCR confirmed the high transcription level of dsHMGR in transgenic cotton lines. The HMGR expression both in transcription and translation level was significantly downregulated in cotton bollworms (helicoverpa armigera) larvae after feeding on the leaves of HMGR transgenic plants. The transcription level of HMGR gene in larvae reared on transgenic cotton leaves was as much as 80.68% lower than that of wild type. In addition, the relative expression level of vitellogenin (Vg, crucial source of nourishment for offspring embryo development) gene was also reduced by 76.86% when the insect larvae were fed with transgenic leaves. The result of insect bioassays showed that the transgenic plant harboring dsHMGR not only inhibited net weight gain but also delayed the growth of cotton bollworm larvae. Taken together, transgenic cotton plant expressing dsRNAs successfully downregulated HMGR gene and impaired the development and survival of target insect, which provided more option for plant pest control.
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http://dx.doi.org/10.7150/ijbs.12463DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4582153PMC
September 2016

Transcriptome characterization analysis of Bactrocera minax and new insights into its pupal diapause development with gene expression analysis.

Int J Biol Sci 2014 12;10(9):1051-63. Epub 2014 Sep 12.

1. Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.

Bactrocera minax is a major citrus pest distributed in China, Bhutan and India. The long pupal diapause duration of this fly is a major bottleneck for artificial rearing and underlying mechanisms remain unknown. Genetic information on B. minax transcriptome and gene expression profiles are needed to understand its pupal diapause. High-throughput RNA-seq technology was used to characterize the B. minax transcriptome and to identify differentially expressed genes during pupal diapause development. A total number of 52,519,948 reads were generated and assembled into 47,217 unigenes. 26,843 unigenes matched to proteins in the NCBI database using the BLAST search. Four digital gene expression (DGE) libraries were constructed for pupae at early diapause, late diapause, post-diapause and diapause terminated developmental status. 4,355 unigenes showing the differences expressed across four libraries revealed major shifts in cellular functions of cell proliferation, protein processing and export, metabolism and stress response in pupal diapause. When diapause was terminated by 20-hydroxyecdysone (20E), many genes involved in ribosome and metabolism were differentially expressed which may mediate diapause transition. The gene sets involved in protein and energy metabolisms varied throughout early-, late- and post-diapause. A total of 15 genes were selected to verify the DGE results through quantitative real-time PCR (qRT-PCR); qRT-PCR expression levels strongly correlated with the DGE data. The results provided the extensive sequence resources available for B. minax and increased our knowledge on its pupal diapause development and they shed new light on the possible mechanisms involved in pupal diapause in this species.
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http://dx.doi.org/10.7150/ijbs.9438DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4183925PMC
May 2015

RNAi silencing of the HaHMG-CoA reductase gene inhibits oviposition in the Helicoverpa armigera cotton bollworm.

PLoS One 2013 2;8(7):e67732. Epub 2013 Jul 2.

Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control, Plant Science & Technology College, Huazhong Agricultural University, Wuhan, China.

RNA interference (RNAi) has considerable promise for developing novel pest control techniques, especially because of the threat of the development of resistance against current strategies. For this purpose, the key is to select pest control genes with the greatest potential for developing effective pest control treatments. The present study demonstrated that the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase; HMGR) gene is a potential target for insect control using RNAi. HMGR is a key enzyme in the mevalonate pathway in insects. A complete cDNA encoding full length HMGR (encoding an 837-aa protein) was cloned from Helicoverpa armigera (Lepidoptera: Noctuidae). The HaHMGR (H. armigera HMGR) knockdown using systemic RNAi in vivo inhibited the fecundity of the females, effectively inhibited ovipostion, and significantly reduced vitellogenin (Vg) mRNA levels. Moreover, the oviposition rate of the female moths was reduced by 98% by silencing HaHMGR compared to the control groups. One-pair experiments showed that both the proportions of valid mating and fecundity were zero. Furthermore, the HaHMGR-silenced females failed to lay eggs (approximate 99% decrease in oviposition) in the semi-field cage performance. The present study demonstrated the potential implications for developing novel pest management strategies using HaHMGR RNAi in the control of H. armigera and other insect pests.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0067732PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3699641PMC
January 2014

Transcriptome profiling of Chironomus kiinensis under phenol stress using Solexa sequencing technology.

PLoS One 2013 20;8(3):e58914. Epub 2013 Mar 20.

Department of Forest Protection, Northeast Forestry University, Harbin, China.

Phenol is a major pollutant in aquatic ecosystems due to its chemical stability, water solubility and environmental mobility. To date, little is known about the molecular modifications of invertebrates under phenol stress. In the present study, we used Solexa sequencing technology to investigate the transcriptome and differentially expressed genes (DEGs) of midges (Chironomus kiinensis) in response to phenol stress. A total of 51,518,972 and 51,150,832 clean reads in the phenol-treated and control libraries, respectively, were obtained and assembled into 51,014 non-redundant (Nr) consensus sequences. A total of 6,032 unigenes were classified by Gene Ontology (GO), and 18,366 unigenes were categorized into 238 Kyoto Encyclopedia of Genes and Genomes (KEGG) categories. These genes included representatives from almost all functional categories. A total of 10,724 differentially expressed genes (P value <0.05) were detected in a comparative analysis of the expression profiles between phenol-treated and control C. kiinensis including 8,390 upregulated and 2,334 downregulated genes. The expression levels of 20 differentially expressed genes were confirmed by real-time RT-PCR, and the trends in gene expression that were observed matched the Solexa expression profiles, although the magnitude of the variations was different. Through pathway enrichment analysis, significantly enriched pathways were identified for the DEGs, including metabolic pathways, aryl hydrocarbon receptor (AhR), pancreatic secretion and neuroactive ligand-receptor interaction pathways, which may be associated with the phenol responses of C. kiinensis. Using Solexa sequencing technology, we identified several groups of key candidate genes as well as important biological pathways involved in the molecular modifications of chironomids under phenol stress.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0058914PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3604134PMC
September 2013

Expression of pheromone biosynthesis activating neuropeptide and its receptor (PBANR) mRNA in adult female Spodoptera exigua (Lepidoptera: Noctuidae).

Arch Insect Biochem Physiol 2010 Sep;75(1):13-27

State Key Laboratory for Plant Diseases and Insect Pests, Institute of Plant Protection Chinese Academy of Agricultural Sciences, Beijing, China.

The full-length cDNA of pheromone biosynthesis activating neuropeptide receptor (PBANR) was cloned from the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae); it included an open reading frame of 1,053 bp encoding 350 amino acids. The PBANR of S. exigua (SePBANR) was structurally characteristic of G protein-coupled receptor and its amino acid sequence shared 98% identity with the PBANR of Spodoptera littoralis. Both pheromone biosynthesis activating neuropeptide (PBAN) and PBANR mRNA abundance were measured in the brain-subesophageal ganglion complex, pheromone gland, ventral nerve cord, and ovary of S. exigua female moths by real-time RT-PCR. The abundance of PBAN mRNA in brain-subesophageal ganglion complex and PBANR mRNA in pheromone gland was significantly greater compared to other tissues, suggesting that the ligand-receptor relationship of PBAN and PBANR exists quantitatively in S. exigua. Both PBAN and PBANR expression displayed a remarkable diurnal rhythm, for they were low and stable during the photophase (07:00-21:00) and increased markedly during the scotophase (with a maximum abundance at 23:30) in 3-day-old female moths. The abundance of PBAN and PBANR increased steadily from the 1st day to the 5th day of the adult female life. The pattern of both diurnal and daily expression of PBAN and PBANR mRNA were coincident with enhanced capacity of sex pheromone release and mating of S. exigua moths during the same period. We infer from these results that pheromone biosynthesis and release in S. exigua is regulated by PBAN via up-regulating synthesis.
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http://dx.doi.org/10.1002/arch.20379DOI Listing
September 2010

Analysis of pupal head proteome and its alteration in diapausing pupae of Helicoverpa armigera.

J Insect Physiol 2010 Mar 30;56(3):247-52. Epub 2009 Oct 30.

Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science & Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China.

The proteomic approach has proven to be an useful tool for understanding insect diapause processes. Using 2D gel electrophoresis and matrix assisted laser/desorption ionization (MALDI) time of flight (TOF), we identified 24 proteins in the head of Helicoverpa armigera pupae with diverse functional characteristics, including cytoskeleton proteins, heat-shock proteins, insect development regulation factors, ATPases, proteins regulating signal pathway and enzymes involved in metabolism, etc. A proteomic comparison between nondiapausing and diapausing pupae revealed three proteins that were present only in nondiapausing pupae, and six proteins represented >or=2.0-fold or
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http://dx.doi.org/10.1016/j.jinsphys.2009.10.008DOI Listing
March 2010

[Influence of temperature and light on the growth and development of Tenodera angustipennis and related preying functional responses].

Ying Yong Sheng Tai Xue Bao 2004 Aug;15(8):1423-6

Institute of Insect Resources, Huazhong Agricultural University, Wuhan, China.

Tenodera angustipennis was raised under laboratory conditions to study the influence of temperature and light on the growth and development of its nymphs. The functional responses of T. angustipennis to Plutella xylostella (L.) larvae and adults and to Lipaphis erysimi (Kaltenbach) were also studied. The results showed that temperature remarkably affected the growth and development of T. angustipennis, but light did not. The Holling models could well describe the functional responses of T. angustipennis to P. xylostella and L. erysimi.
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August 2004