Knock-in of exogenous sequences based on CRISPR/Cas9 targeting autosomal genes and sex chromosomes in the diamondback moth,Plutella xylostella

Genetic pest control strategies based on precise sex separation and only releasing sterile males can be accomplished by site-specific genome editing.In the current study,we showed that the mutation of single-allele Pxfl(2)d can significantly impair the normal mating behavior and testis development in male adults of the notorious cruciferous insect pest Plutella xylostella,in addition to its known functions in the ovarian development in female adults and egg hatching.Subsequent CRISPR/Cas9-based knock-in experiments revealed that site-specific integration of an exogenous green fluorescent protein(GFP)gene into autosomal Pxfl(2)d for labelling mutants could be achieved.However,this gene is not a suitable target for GFP insertion to establish a genetically stable knock-in strain because of the severe decline in reproductive capacity.We further screened for the W-chromosome-linked and Z-chromosome-linked regions to test the knock-in efficiency mediated by CRISPR/Cas9.The results verified that both types of chromosomes can be targeted for the site-specific insertion of exogenous sequences.We ultimately obtained a homozygous knock-in strain with the integration of both Cas9 and cyan fluorescent protein(CFP)expression cassettes on a Z-linked region in P.xylostella,which can also be used for early sex detection.By injecting the sgRNA targeting Pxfl(2)d alone into the eggs laid by female adults of the Z-Cas9-CFP strain,the gene editing efficiency reached 29.73%,confirming the success of expressing a functional Cas9 gene.Taken together,we demonstrated the feasibility of the knock-in of an exogenous gene to different genomic regions in P.xylostella,while the establishment of a heritable strain required the positioning of appropriate sites.This study provides an important working basis and technical support for further developing genetic strategies for insect pest control.

Influence of genes,sex,age and environment on the onset of autoimmune hepatitis

The pathogenesis of autoimmune hepatitis (AIH) is complex. However,it is believed that a susceptible individual,owing to his genetic background,sex and age,can develop the disease following exposure to an environmental trigger. Autoimmune hepatitis does not follow a Mendelian pattern of inheritance; hence no single causative genetic locus has been identified. However,several genes,inside and outside the HLA locus,have been linked to an increased susceptibility to AIH. Epidemiological evidence also suggests that the sex and age of the patient plays a role in AIH pathogenesis as the disease onset occurs mainly in the two first decades of life and a higher disease incidence is observed in females. No environmental trigger has been identified,but several have been proposed,mainly viruses and xenobiotics. This article aims at reviewing the current knowledge on susceptibility factors leading to AIH and putative triggers,emphasizing fundamental mechanisms responsible for the break of liver immunological tolerance.

Analysis of the Offspring Sex Ratio of Chicken by Using Molecular Sexing

The overall sex ratio of offspring （dead embryos and hatch chicks） from all the fertilized eggs of 140 hens collected for 30 days was studied using duplex PCR of certain fragments of sex chromosomes. Additional 894 dead embryos over a period of 21 days of incubation were also investigated to verify the sex ratio of the dead embryos. The sex of the early dead embryos was identified using this molecular sexing technique. The sex ratio of the hatch chicks and the total offspring of the hens investigated in this experiment did not differ from the expected sex ratio （i.e., 1：1）., However, the number of female dead embryos was significantly more than that of males. The data indicated that the different physiologic function of males and females contributed to female-biased mortality during incubation. It was also found by further analysis that the sex ratios of the offspring of some hens were significantly biased to female or male over the period investigated, which suggested that the sex ratio of offspring might be influenced by the maternal condition to some degrees.

高职资源勘查类专业Auto CAD课程产学融合教学模式探索与构建

以资源勘查类专业的Auto CAD课程教学为例,探讨如何有效地进行教学管理和完成教学设计,以适应产学融合、培养高质量的技术型人才的需求。通过针对性的课程设计,融入资源勘查类专业的知识点并引导学生运用知识点,能在一定程度上提高学生的学习兴趣,有利于实现以需求为导向的教学目标,使产学互动无缝衔接。

Digital CEOs in Digital Enterprises:Automating,Augmenting,and Parallel in Metaverse/CPSS/TAOs

BIG models or foundation models are rapidly emerging as a key force in advancing intelligent societies[1]–[3]Their significance stems not only from their exceptional ability to process complex data and simulate advanced cognitive functions,but also from their potential to drive innovation across various industries.

Forecasting risk using auto regressive integrated moving average approach: an evidence from S&P BSE Sensex

The primary objective of the paper is to forecast the beta values of companies listed on Sensex,Bombay Stock Exchange(BSE).The BSE Sensex constitutes 30 top most companies listed which are popularly known as blue-chip companies.To reach out the predefined objectives of the research,Auto Regressive Integrated Moving Average method is used to forecast the future risk and returns for 10 years of historical data from April 2007 to March 2017.Validation accomplished by comparison of forecasted and actual beta values for the hold back period of 2 years.Root-Mean-Square-Error and Mean-Absolute-Error both are used for accuracy measurement.The results revealed that out of 30 listed companies in the BSE Sensex,10 companies’exhibits high beta values,12 companies are with moderate and 8 companies are with low beta values.Further,it is to note that Housing Development Finance Corporation(HDFC)exhibits more inconsistency in terms of beta values though the average beta value is lowest among the companies under the study.A mixed trend is found in forecasted beta values of the BSE Sensex.In this analysis,all the p-values are less than the F-stat values except the case of Tata Steel and Wipro.Therefore,the null hypotheses were rejected leaving Tata Steel and Wipro.The values of actual and forecasted values are showing the almost same results with low error percentage.Therefore,it is concluded from the study that the estimation ARIMA could be acceptable,and forecasted beta values are accurate.So far,there are many studies on ARIMA model to forecast the returns of the stocks based on their historical data.But,hardly there are very few studies which attempt to forecast the returns on the basis of their beta values.Certainly,the attempt so made is a novel approach which has linked risk directly with return.On the basis of the present study,authors try to through light on investment decisions by linking it with beta values of respective stocks.Further,the outcomes of the present study undoubtedly useful to academicians,researchers,and policy makers in their respective area of studies.

The interventional effect of Polygonatum cyrtonema Hua polysaccharide on atherosclerosis in mice of different sexes

In the present study,we investigated the intervention effects of a purified Polygonatum cyrtonema polysaccharide(PCP)on high-fat diet(HFD)-induced atherosclerosis in male and female LDLr-/-mice.Results showed that HFD caused severe dyslipidemia,atherosclerotic lesions,oxidative damages and inflammation in male and female mice,and these effects seemed to be more pronounced in males than in females.However,the above variations could be dose-dependently reversed by PCP treatment,and the intervention effects on males were greater than those on females.Nuclear factor kappa-B(NF-κB),mitogen-activated protein kinase(MAPKs)and protein kinase B(Akt)are 3 pivotal signaling pathways mediating the development of atherosclerosis.Consistently,PCP was also found to significantly decrease the phosphorylation of p65,p38,extracellular-regulated kinase 1/2(ERK1/2)and Akt,and increase the protein expression of inhibitor of NF-κB(IκB)in the aortas of male and female mice induced by HFD.Taken together,these findings indicated that PCP could be effective for the prevention of atherosclerosis,and the intervention effect of PCP on male mice was more obvious than that of female mice.

Hepatoprotective effects of Xiaoyao San formula on hepatic steatosis and inflammation via regulating the sex hormones metabolism

BACKGROUND The modified Xiaoyao San(MXS)formula is an adjuvant drug recommended by the National Health Commission of China for the treatment of liver cancer,which has the effect of preventing postoperative recurrence and metastasis of hepatocellular carcinoma and prolonging patient survival.However,the molecular mechanisms underlying that remain unclear.AIM To investigate the role and mechanisms of MXS in ameliorating hepatic injury,steatosis and inflammation.METHODS A choline-deficient/high-fat diet-induced rat nonalcoholic steatohepatitis(NASH)model was used to examine the effects of MXS on lipid accumulation in primary hepatocytes.Liver tissues were collected for western blotting and immunohisto chemistry(IHC)assays.Lipid accumulation and hepatic fibrosis were detected using oil red staining and Sirius red staining.The serum samples were collected for biochemical assays and NMR-based metabonomics analysis.The inflammation/lipid metabolism-related signaling and regulators in liver tissues were also detected to reveal the molecular mechanisms of MXS against NASH.RESULTS MXS showed a significant decrease in lipid accumulation and inflammatory response in hepatocytes under metabolic stress.The western blotting and IHC results indicated that MXS activated AMPK pathway but inhibited the expression of key regulators related to lipid accumulation,inflammation and hepatic fibrosis in the pathogenesis of NASH.The metabonomics analysis systemically indicated that the arachidonic acid metabolism and steroid hormone synthesis are the two main target metabolic pathways for MXS to ameliorate liver inflammation and hepatic steatosis.Mechanistically,we found that MXS protected against NASH by attenuating the sex hormone-related metabolism,especially the metabolism of male hormones.CONCLUSION MXS ameliorates inflammation and hepatic steatosis of NASH by inhibiting the metabolism of male hormones.Targeting male hormone related metabolic pathways may be the potential therapeutic approach for NASH.

Sex and racial disparities in non-alcoholic fatty liver disease-related cardiovascular events: National inpatient sample analysis (2019)

BACKGROUND Non-alcoholic fatty liver disease(NAFLD)increases cardiovascular disease(CVD)risk irrespective of other risk factors.However,large-scale cardiovascular sex and race differences are poorly understood.AIM To investigate the relationship between NAFLD and major cardiovascular and cerebrovascular events(MACCE)in subgroups using a nationally representative United States inpatient sample.METHODS We examined National Inpatient Sample(2019)to identify adult hospitalizations with NAFLD by age,sex,and race using ICD-10-CM codes.Clinical and demographic characteristics,comorbidities,and MACCE-related mortality,acute myocardial infarction(AMI),cardiac arrest,and stroke were compared in NAFLD cohorts by sex and race.Multivariable regression analyses were adjusted for sociodemographic characteristics,hospitalization features,and comorbidities.RESULTS We examined 409130 hospitalizations[median 55(IQR 43-66)years]with NFALD.NAFLD was more common in females(1.2%),Hispanics(2%),and Native Americans(1.9%)than whites.Females often reported non-elective admissions,Medicare enrolment,the median age of 55(IQR 42-67),and poor income.Females had higher obesity and uncomplicated diabetes but lower hypertension,hyperlipidemia,and complicated diabetes than males.Hispanics had a median age of 48(IQR 37-60),were Medicaid enrollees,and had non-elective admissions.Hispanics had greater diabetes and obesity rates than whites but lower hypertension and hyperlipidemia.MACCE,all-cause mortality,AMI,cardiac arrest,and stroke were all greater in elderly individuals(P<0.001).MACCE,AMI,and cardiac arrest were more common in men(P<0.001).Native Americans(aOR 1.64)and Asian Pacific Islanders(aOR 1.18)had higher all-cause death risks than whites.CONCLUSION Increasing age and male sex link NAFLD with adverse MACCE outcomes;Native Americans and Asian Pacific Islanders face higher mortality,highlighting a need for tailored interventions and care.

Effects of heterotopic autotransplantation of cryopreserved-thawed ovarian tissues on sex hormone level

Objective: The aim of our study was to measure and compare the serum hormone level of transplant group with blank control and castrated control groups after heterotopic autotransplantation of cryopreserved-thawed ovarian tissues into back muscles. Methods: A total of 40 SPF-SD female rats(5–6 week-old) were randomly divided into three groups: blank control group(group A), castration control group(group B) and transplant group(group C). Ovaries were removed by surgical procedure, then after cryopreservation and thawing procedures the ovarian tissues were implanted into the back muscles of mice in group C. After 4 weeks of ovarian tissues transplantation, all rats blood sampling were measured for E2, LH and FSH hormone levels by ELISA. Results: E2 level was significantly higher in group C and group A than group B [(38.98 ± 5.66) pg/mL,(8.14 ± 3.24) pg/mL; P < 0.05) and [(36.30 ± 6.90) pg/mL,(8.14 ± 3.24) pg/mL; P < 0.05)]. However, E2 level in group C and group A had no significant difference. FSH level in group B, group A and group C was(18.87 ± 2.54) nmol/L,(7.77 ± 0.87) nmol/L and(9.39 ± 2.12) nmol/L respectively. FSH level increased significantly in group B compared with group A, and the difference had statistical significance(P < 0.05). FSH level was slightly increased in group C compared with group A, and the difference was not statistically significant(P > 0.05), but compared with group B, FSH level was significantly reduced and being statistically significant(P < 0.05). Conclusion: Autotransplantation of cryopreserved-thawed ovarian tissue into back muscles can sustain follicular development and re-establish endogenous hormone production by restoring the factors such as angiogenesis and innervations at the graft site.

Trends in in ovo sexing technologies:insights and interpretation from papers and patents

Numerous researchers and institutions have been developing in ovo sexing technologies to improve animal welfare by identifying male embryos in an early embryonic stage and disposing of them before pain perception.This review gives a complete overview of the technological approaches reported in papers and patents by performing a thorough search using Web of Science and Patstat/Espacenet databases for papers and patents,respectively.Based on a total of 49 papers and 115 patent families reported until May 2023 worldwide,11 technology categories were defined:6 non-optical and 5 optical techniques.Every category was described for its characteristics while assessing its potential for application.Next,the dynamics of the publications of in ovo sexing techniques in both paper and patent fields were described through growth curves,and the interest or actual status was visualized using the number of paper citations and the actual legal status of the patents.When comparing the reported technologies in papers to those in patents,scientific gaps were observed,as some of the patented technologies were not reported in the scientific literature,e.g.,ion mobility and mass spectrometry approaches.Generally,more diverse approaches in all categories were found in patents,although they do require more scientific evidence through papers or industrial adoption to prove their robustness.Moreover,although there is a recent trend for non-invasive techniques,invasive methods like analyzing DNA through PCR or hormones through immunosensing are still being reported(and might continue to be)in papers and patents.It was also observed that none of the technologies complies with all the industry requirements,although 5 companies already entered the market.On the one hand,more research and harmony between consumers,industry,and governments is necessary.On the other hand,close monitoring of the market performance of the currently available techniques will offer valuable insights into the potential and expectations of in ovo sexing techniques in the poultry industry.

Sex Determination in Homo sapiens as a Multi-Step Process: Potential for Development of Variants and Sex Differences in Disease Risk

Reproduction via cis-binary mechanisms appears to have evolved fairly early in the evolution of complex organisms, and a system committed to prior to evolution of humans. While the evolution of a chromosomal-specific approach has been a successful strategy for survival of a large variety of species including humans, the fidelity of sex determination leading to 100% cis-binary outcomes is not achieved in many species, with evidence for homosexual or bisexual behaviour evident in more than 1500 species. Thus, such outcomes indicates that sex determination is a multi-step process and not a single event, and as such, could lead to the appearance of variants during the process which developed much earlier than humans. Variants could arise either due to intrinsic variation in the steps of determination, or also be influenced by environmental factors of a biological or psychological nature. In contrast to homosexual variants which do not require interventions such as hormone therapy or surgery, expression of gender dysphoria, is more based in psychology, but also has biological underpinnings and can be influenced by such hormonal interventions and surgery. While the numbers of those with gender dysphoria is small (~0.6% - 1.0% of population), the attention given to this issue raises the possibility of biological and psychological environmental factors impacting the emergence of some of those expressing gender dysphoria. Furthermore, transitioning from male-to-female or female-to-male can have consequences regarding disease risks latter in life, including the appearance of autoimmune diseases. This review will attempt to review some of the evidence regarding sex determination, discuss why the system has potentially not been improved upon during evolution, how a potential role for sex chromosome function on neurodevelopment may be central to variation in humans, and how commitment to the current strategy is likely integrated into other sex-related events such as puberty, pregnancy, and menopause to ensure species survival. It will also discuss how variants in sex determination could contribute to sex differences in disease risk and how epigenetic modifications could play a role in such risk. .

Overexpression of Kdm6b induces testicular differentiation in a temperature-dependent sex determination system

In reptiles,such as the red-eared slider turtle(Trachemys scripta elegans),gonadal sex determination is highly dependent on the environmental temperature during embryonic stages.This complex process,which leads to differentiation into either testes or ovaries,is governed by the finely tuned expression of upstream genes,notably the testis-promoting gene Dmrt1 and the ovary-promoting gene Foxl2.Recent studies have identified epigenetic regulation as a crucial factor in testis development,with the H3K27me3 demethylase KDM6B being essential for Dmrt1 expression in T.s.elegans.However,whether KDM6B alone can induce testicular differentiation remains unclear.In this study,we found that overexpression of Kdm6b in T.s.elegans embryos induced the male development pathway,accompanied by a rapid increase in the gonadal expression of Dmrt1 at 31°C,a temperature typically resulting in female development.Notably,this sex reversal could be entirely rescued by Dmrt1 knockdown.These findings demonstrate that Kdm6b is sufficient for commitment to the male pathway,underscoring its role as a critical epigenetic regulator in the sex determination of the red-eared slider turtle.

Influence of sex on outcomes of liver transplantation for hepatocellular carcinoma:a multicenter cohort study in China

Objective:Sex-specific differences are observed in various liver diseases,but the influence of sex on the outcomes of hepatocellular carcinoma(HCC)after liver transplantation(LT)remains to be determined.This study is the first Chinese nationwide investigation of the role of sex in post-LT outcomes in patients with HCC.Methods:Data for recipients with HCC registered in the China Liver Transplant Registry between January 2015 and December 2020 were analyzed.The associations between donor,recipient,or donor-recipient transplant patterns by sex and the post-LT outcomes were studied with propensity score matching(PSM).The survival associated with different sex-based donor-recipient transplant patterns was further studied.Results:Among 3,769 patients enrolled in this study,the 1-,3-,and 5-year overall survival(OS)rates of patients with HCC after LT were 96.1%,86.4%,and 78.5%,respectively,in female recipients,and 95.8%,79.0%,and 70.7%,respectively,in male recipients after PSM(P=0.009).However,the OS was comparable between recipients with female donors and male donors.Multivariate analysis indicated that male recipient sex was a risk factor for post-LT survival(HR=1.381,P=0.046).Among the donor-recipient transplant patterns,the male-male donor-recipient transplant pattern was associated with the poorest post-LT survival(P<0.05).Conclusions:Our findings highlighted that the post-LT outcomes of female recipients were significantly superior to those of male recipients,and the male-male donor-recipient transplant pattern was associated with the poorest post-LT survival.Livers from male donors may provide the most benefit to female recipients.Our results indicate that sex should be considered as a critical factor in organ allocation.

基于智慧职教平台的混合式教学模式课程设计与教学实践研究——以Auto CAD绘图课程为例

传统Auto CAD绘图课程的教学模式是老师不断输出知识和操作演示,学生被动接受和模仿学习,学生未能真正将知识进行内化,也没有思考后的输出和应用。该方式教学内容与企业需求脱轨,学生只能完成课堂布置的练习及课后作业,缺乏学习探索的主动性。据此,将现代信息化教学手段与普通线下教学方式进行糅合,并在此基础上进行课程内容重构、教学过程设计和多元化评价体系等,以求达到更好的教学效果,培养出企业需要、社会满意的学生。

Epidemiological Profile of Men Who Have Sex with Men: First Data about Male Sex Workers in Central African Republic (CAR)

Objectives: Sex work is not well documented among African men. The aim of this study was to describe the epidemiological profile of men who have sex with men (MSM) and eventually to determine the proportion of sex workers (SW) among them, as well as the proportion of MSM who have been victims of gender-based violence (GBV). Material and Methods: A cross-sectional study coordinated by the Direction de la Médecine du Travail was carried out at the headquarters of the association ALTERNATIVES in Bangui Bangui from July 1 to October 31, 2021. Consenting MSM present during the study period were systematically included. Sociodemographic variables, those relating to the future vision of the activity and to GBV were collected and analyzed using Epi-Info version 7 software. Results: Forty MSM with an average age of 23 years and mainly secondary education (75.0%) were included. The vast majority of MSM were unemployed (85.0%). 45.0% had been victims of GBV and wanted to stop working as an MSM (47.5%), and almost 2/3 (65.0%) would accept another income-generating activity in exchange for the MSM. Violence was sexual (32.5%), economic (22.5%), physical (20.0%), verbal (12.5%) and psychological (12.5%). Conclusion: The MSM were mainly young, poorly educated, unemployed, and almost half were victims of GBV. The desire to change MSM activity to another income-generating activity alongside that of MSM shows that many of them are SW, workers in the informal sector. This must be taken into account in prevention activities, even if the data needs to be confirmed on a much larger sample.

Hard life for sons in the nest?Sex-dependent offspring mortality in Great Tits in urban and forest areas

Sex-biased mortality can occur in birds during development,for example due to sexual differences in energy requirement and/or environmental sensitivity,or the effects of sex hormones or sex differences in the expression of mutations linked to sex chromosomes.The extent of sex-bias in mortality may also be related to environmental conditions that influence offspring development and survival.Urban areas often provide poorer conditions for nestling development resulting in higher offspring mortality compared to natural areas,which may accelerate sex differences in offspring mortality in cities.To test this hypothesis,we examined the sex ratio of dead offspring in Great Tits(Parus major),using 427 samples of unhatched eggs and dead nestlings collected in two urban and two forest sites between 2013 and 2019.The ratio of males in the whole sample of dead offspring(56.9%)was significantly higher than expected by an 1:1 ratio,and the strongest sex biases were detected in urban areas(57.6%males)and in young nestlings(<14 days old,59.0%males).However,the sex ratios of dead offspring did not differ significantly among study sites and between offspring developmental stages.29.3%of unhatched eggs contained a visible embryo,and the proportion of embryo-containing unhatched eggs did not differ significantly between urban and forest study sites.These results suggest male-biased offspring mortality in Great Tits,and highlight the need of large datasets to detect subtle differences between habitats and developmental stages.

Changes in sexual activity in young women during pregnancy

Objective:To conduct changes in sexual activity during pregnancy and its related factors in pregnant women.Methods:The present descriptive cross-sectional study was conducted in 2017 on pregnant women who referred to the women's clinic of Afzalipour Hospital in the southeast of Iran.Participants were included in the study through convenient sampling.The data collection tool was a researcher-made questionnaire consisting of two parts of personal social information of the couple and questions to measure the level of sexual activity and the attitude of the subjects during pregnancy compared to before pregnancy.Results:201 Pregnant women were included.The average age of pregnant women was(27.3±6.1)years and their average gestational age was(24.7±11.8)years.More than 62%of women had decreased sexual activity.There was a significant relationship between the amount of changes in sexual activity and delivery time(P=0.013),abortion history(P=0.001)and premature birth history(P=0.002).Most pregnant women believed that sex during pregnancy caused damage to the fetus(67.7%).A decrease in the intensity of sexual desire was reported in 63%of pregnant women.More than 60%of the subjects did not consult with doctors and midwives with regards to sexual issues(63.5%).The most common reason for not consulting was not feeling the need(32%).Most women experienced back pain during(42.8%)and after(39.8%)intercourse.Conclusions:Changes in sex life during pregnancy are often caused by the lack of sexual knowledge and the increase in misconceptions among couples,which can affect the quality of relationships.The role of education is essential.Therefore,it is suggested that by including sexual counseling along with pregnancy care,wrong beliefs and information among women will be corrected.

Associated Factors with Early Sexual Intercourse among In-School Adolescents in Brazzaville

Background: Early sexual intercourse (ESI) is a global public health problem. In Congo, associated factors with ESI have been under-studied. Objective: To analyse factors associated with early sexual intercourse among in-school adolescents in Brazzaville. Method: A cross-sectional analytic study with three-stage random sampling was carried out from March 2 to June 2, 2022 in Brazzaville secondary and high schools. Adolescents in the selected schools were interviewed about their sexual and social behaviour, and their family and school environment. Multivariate analysis using logistic regression was used to identify factors associated with ESI. The significance level of the p-value was set at 5% and the confidence interval at 95%. Results: Of a total of 1100 adolescents surveyed, 800 (72.7%) had ESI, 98% of them were female. The main reason for early initiation of sexual intercourse was curiosity (81.8%). Non-use of condoms and multiple sexual partnerships were noted respectively in 98.7% and 68.7% of adolescents who had experienced ESI. Female gender (p Conclusion: The prevalence of ESI is high, and the factors identified in this study may help to reduce it.

The Ca^(2+)/CaN/ACC and cAMP/PKA/HK signal pathways are required for PBAN-mediated sex pheromone biosynthesis in Conogethes punctiferalis

Conogethes punctiferalis is a crop and fruit pest that has caused serious economic losses to agricultural production.This pest relies heavily on its sex pheromone to ensure sexual encounters and subsequent mating success.However,the molecular mechanism underlying sex pheromone biosynthesis in this species remains elusive.The present study investigated the detailed mechanism underlying PBAN-regulated sex pheromone biosynthesis in C.punctiferalis by transcriptome sequencing of the C.punctiferalis pheromone glands(PGs)and subsequent functional identification of the target genes.The results showed that female mating started from the first scotophase,and peaked at the second to fifth scotophases in accordance with the release of sex pheromones.PBAN regulated sex pheromone biosynthesis by employing Ca^(2+)and cAMP as secondary messengers,as demonstrated by RNA interference(RNAi),pharmacological inhibitors,and behavioral assays.Further investigation revealed that calcineurin(CaN)and acetyl-CoA carboxylase(ACC)were activated by PBAN/Ca^(2+)signaling,and the RNAimediated knockdown of CaN and ACC transcripts significantly reduced sex pheromone production,ultimately leading to a significantly reduced ability of females to attract males.Importantly,hexokinase(HK)was found to regulate sex pheromone biosynthesis in response to the PBAN/cAMP/PKA signaling pathway,as demonstrated by RNAi,enzyme activity,and pharmacological inhibitor assays.Furthermore,Far2 and Desaturase1 were found to participate in PBAN-regulated sex pheromone biosynthesis.Altogether,our findings revealed that PBAN regulates sex pheromone biosynthesis through the PBANR/Ca^(2+)/CaN/ACC and PBANR/cAMP/PKA/HK pathways in C.punctiferalis,which enriches our comprehension of the details of sex pheromone biosynthesis in moths.