The role of dietary preferences in osteoarthritis: a Mendelian randomization study using genome-wide association analysis data from the UK Biobank.

Background: To understand the impact of individual preferences for specific dietary items on OA, and to help inform the development of effective and targeted OA prevention and management strategies, we performed a Mendelian randomization analysis between dietary preferences and osteoarthritis. Methods: This study utilized genetic data from the UK Biobank to investigate the association between OA and 21 different common dietary items. Instrumental variables representing European populations were carefully selected based on their genetic significance and linkage disequilibrium. In cases where a dietary item had few relevant genetic markers, a more lenient selection threshold was applied. To prevent bias, the analysis excluded single nucleotide polymorphisms (SNPs) associated with factors such as body mass index (BMI) and cholesterol. Using inverse-variance weighting (IVW) and Mendelian randomization, significant associations were detected between certain dietary items and OA. Results: Using Mendelian randomization to examine the relationship between 21 different dietary items and OA, significant associations were found for coffee, peas, watercress, and cheese, where the first two had a promoting effect and the last two an inhibiting effect on OA. Due to heterogeneity in the test results for cheese, a random IVW representation was used. The results of sensitivity analysis showed no significant heterogeneity or horizontal pleiotropy in the selected SNPS, demonstrating the reliability of Mendelian randomization analysis. Conclusion: This study identified coffee, peas, watercress, and cheese as food items that may have significant dietary effects on osteoarthritis. This information may be useful to consider in the development of OA management strategies.

Developing a Machine-Learning Predictive Model for Retention of Posterior Cruciate Ligament in Patients Undergoing Total Knee Arthroplasty.

OBJECTIVE: Predicting whether the posterior cruciate ligament (PCL) should be preserved during total knee arthroplasty (TKA) procedures is a complex task in the preoperative phase. The choice to either retain or excise the PCL has a substantial effect on the surgical outcomes and biomechanical integrity of the knee joint after the operation. To enhance surgeons' ability to predict the removal and retention of the PCL in patients before TKA, we developed machine learning models. We also identified significant feature factors that contribute to accurate predictions during this process. METHODS: Patients' data on TKA continuously performed by a single surgeon who had intended initially to undergo implantation of cruciate-retaining (CR) prostheses was collected. During the sacrifice of PCL, we utilized anterior-stabilized (AS) tibial bearings. The dataset was split into CR and AS categories to form distinct groups. Relevant information regarding age, gender, body mass index (BMI), the affected side, and preoperative diagnosis was extracted by reviewing the medical records of the patients. To ensure the authenticity of the research, an initial step involved capturing X-ray images before the surgery. These images were then analyzed to determine the height of the medial condyle (MMH) and lateral condyle (LMH), as well as the ratios between MLW and MMH and MLW and LMH. Additionally, the insall-salvati index (ISI) was calculated, and the severity of any varus or valgus deformities was assessed. Eight machine-learning methods were developed to predict the retention of PCL in TKA. Risk factor analysis was performed using the SHApley Additive exPlanations method. RESULTS: A total of 307 knee joints from 266 patients were included, among which there were 254 females and 53 males. A stratified random sampling technique was used to split patients in a 70:30 ratio into a training dataset and a testing dataset. Eight machine-learning models were trained using data feeding. Except for the AUC of the LGBM Classifier, which is 0.70, the AUCs of other machine learning models are all lower than 0.70. In importance-based analysis, ISI, MMH, LMH, deformity, and age were confirmed as important predictive factors for PCL retention in operations. CONCLUSION: The LGBM Classifier model achieved the best performance in predicting PCL retention in TKA. Among the potential risk factors, ISI, MMH, LMH, and deformity played essential roles in the prediction of PCL retention.

Mosquito E-20-Monooxygenase Gene Knockout Increases Dengue Virus Replication in Aedes aegypti Cells.

E-20-monooxygenase (E20MO) is an enzymatic product of the shade (shd) locus (cytochrome p450, E20MO). Initially discovered in Drosophila, E20MO facilitates the conversion of ecdysone (E) into 20-hydroxyecdysone (20E) and is crucial for oogenesis. Prior research has implicated 20E in growth, development, and insecticide resistance. However, little attention has been given to the association between the E20MO gene and DENV2 infection. The transcriptome of Ae. aegypti cells (Aag2 cells) infected with DENV2 revealed the presence of the E20MO gene. The subsequent quantification of E20MO gene expression levels in Aag2 cells post-DENV infection was carried out. A CRISPR/Cas9 system was utilized to create an E20MO gene knockout cell line (KO), which was then subjected to DENV infection. Analyses of DENV2 copies in KO and wild-type (WT) cells were conducted at different days post-infection (dpi). Plasmids containing E20MO were constructed and transfected into KO cells, with pre- and post-transfection viral copy comparisons. Gene expression levels of E20MO increased after DENV infection. Subsequently, a successful generation of an E20MO gene knockout cell line and the verification of code-shifting mutations at both DNA and RNA levels were achieved. Furthermore, significantly elevated DENV2 RNA copies were observed in the mid-infection phase for the KO cell line. Viral RNA copies were lower in cells transfected with plasmids containing E20MO, compared to KO cells. Through knockout and plasmid complementation experiments in Aag2 cells, the role of E20MO in controlling DENV2 replication was demonstrated. These findings contribute to our understanding of the intricate biological interactions between mosquitoes and arboviruses.

Application of thifluzamide to stem rot in peppers: Infection and control mechanisms of sclerotium rolfsii.

In recent years, the fungal disease 'pepper stem rot', contracted from the soil-borne pathogen sclerotium rolfsii, has been increasing year by year, causing significant losses to the pepper (Capsicum annuum L.) industry. To investigate the infection mechanism of stem rot, the fungus S. rolfsii was used to infect the roots of pepper plants, and was found to affect root morphology and reduce root activity, which subsequently inhibited root growth and development. With fungal infestation, its secretions (oxalic acid, PG and PMG enzyme) were able to break normal tissues in the stem base and induced the burst of the active oxygen, which leads to injury aggravation. Morphological observations of the site of damage at the base of the stem using SEM revealed that the vascular bundles and stomata were completely blocked by hyphae, resulting in a blockade of material exchange in the plant. It was subsequently found that most of the stomata in the leaves were closed, which caused the leaves to lose their ability to photosynthesize, then turned yellow, wilt, shed, and the plant died. Commercialized fungicide thifluzamide with excellent in vitro (EC50 = 0.1 µg/mL) and in vivo curative (EC50 = 29.2 µg/mL) antifungal activity was selected to control the stem rot disease in peppers. The results demonstrated that it was able to suppress the secretion of associated pathogenic factors and reduce the outbursts of reactive oxygen species, thus reducing the damage caused by S. rolfsii at the base of the plant's stem and also enhancing the root activity of the infected plant, thereby promoting root growth. It could also inhibit fungal growth, unblock the vascular bundles and stomata, maintain a balance of material and energy exchange within the plant, and thus restore the damaged plant to its normal growth capacity. All the results will provide an adequate reference for the prevention and control of stem rot disease on peppers with thifluzamide.

The effect of artificial light at night (ALAN) on the characteristics of diapause of Aedes albopictus.

BACKGROUND: Recently, the effect of artificial light at night (ALAN) on the physiology and behavior of insects has gradually attracted the attention of researchers and has become a new research topic. Aedes albopictus is an important vector that poses a great public health risk. Further studies on the diapause of Ae. albopictus can provide a basis for new vector control, and it is also worth exploring whether the effect of ALAN on the diapause of Ae. albopictus will provide a reference for the prevention and control of infectious diseases mediated by Ae. albopictus. METHODS: In this study, we experimentally studied the diapause characteristics of different geographical strains of Ae. albopictus under the interference of ALAN, explored the effect of ALAN on the diapause of Ae. albopictus and explored the molecular mechanism of ALAN on the diapause process through RNA-seq. RESULTS: As seen from the diapause incidence, Ae. albopictus of the same geographic strain showed a lower diapause incidence when exposed to ALAN. The differentially expressed genes (DEGs) were mainly enriched in signaling and metabolism-related pathways in the parental females and diapause eggs of the ALAN group. CONCLUSIONS: ALAN inhibits Ae. albopictus diapause. In the short photoperiod induced diapause of Ae. albopictus in temperate strain Beijing and subtropical strain Guangzhou, the disturbance of ALAN reduced the egg diapause rate and increased the egg hatching rate of Ae. albopictus, and the disturbance of ALAN also shortened the life cycle of Ae. albopictus eggs after hatching.

Identification and Genetic Diversity Analysis of the Pathogen of Anthracnose of Pepper in Guizhou.

Anthracnose of pepper is a significant disease caused by Colletotrichum spp. In 2017 and 2021, 296 isolates were obtained from 69 disease samples. Through morphological analysis, pathogenicity detection, and polygenic phylogenetic analysis, the above strains were attributed to 10 species: C. scovillei, C. fructicola, C. karstii, C. truncatum, C. gloeosporioides, C. kahawae, C. boninense, C. nymphaeae, C. plurivorum, and C. nigrum. C. scovillei had the most strains (150), accounting for 51.02% of the total isolates; C. fructicola came in second (72 isolates), accounting for 24.49%. Regarding regional distribution, Zunyi City has the highest concentration of strains-92 strains total, or 34.18%-across seven species. Notably, this investigation showed that C. nymphaeae infected pepper fruit for the first time in China. Genetic diversity analysis showed that C. fructicola could be divided into seven haplotypes, and the population in each region had apparent genetic differentiation. However, the genetic distance between each population was not significantly related to geographical distance. Neutral detection and nucleotide mismatch analysis showed that C. fructicola might have undergone population expansion.

Insecticide resistance status of Aedes aegypti in border areas of Yunnan Province.

BACKGROUND: Aedes aegypti is a main vector of arboviral diseases, principally dengue, chikungunya, and Zika. Insecticides remain the most effective vector control method. Pyrethroid is the main insecticide currently used, and the long-term use of insecticides can cause mosquitoes to develop knockdown resistance. Studying the mutation sites and genotypes of Ae. aegypti can reveal the mutation characteristics and regional distribution of the kdr gene in an Ae. aegypti population. Testing for a correlation between the mutation rate in various populations and pyrethrin resistance can clarify the resistance mechanism. RESULTS: The bioassay results showed that all 15 populations are resistant. In the study of the kdr gene, three non-synonymous mutations were identified in the DNA of first generation females from the wild Ae. aegypti population: S989P (TCC-CCC), V1016G (GTA-GGA), and F1534C (TTC-TGC). The mortality rate of the various populations was correlated with the mutation rate at the V1016G + F1534C locus, but not the S989P + V1016G locus. CONCLUSION: Aedes aegypti populations in border regions of Yunnan Province are resistant to permethrin and beta-cyfluthrin. The insecticidal effect of beta-cyfluthrin is stronger than that of permethrin. The mutation rate at sites V1016G + F1534C is negatively correlated with the mortality of Ae. aegypti based on bioassays. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Aedes aegypti Beta-1,3-Glucan-Binding Protein Inhibits Dengue and ZIKA Virus Replication.

GNBPB6, a beta-1,3-glucan-binding protein, was identified in the transcriptome of Aedes aegypti (A. aegypti) with dengue (DENV), Zika (ZIKV), and chikungunya viruses (CHIKV). In this study, we not only clarified that DENV2 and ZIKV regulate the changes in GNBPB6 expression but also identified the relationship of this gene with viral infections. The changes in GNBPB6 expression were quantified and showed a decrease in A. aegypti cells (Aag2 cells) at 2 dpi and 3 dpi and an increase at 4 dpi and 5 dpi (p < 0.05). A significant increase was observed only at 5 dpi after DENV2 infection. Subsequently, a GNBPB6 knockout (KO) cell line was constructed using the CRISPR/Cas9 system, and the DENV2 and ZIKV RNA copies, along with cell densities, were quantified and compared between the KO and wild type (WT) cells at different dpi. The result showed that DENV2 and ZIKV RNA copies were significantly increased in the KO cell line with no significant change in cell growth. Finally, DENV2 copies decreased after GNBPB6 was complemented in the KO. In conclusion, GNBPB6 knockout and complementation in Aag2 cells revealed that GNBPB6 can inhibit the replication of both DENV2 and ZIKV. These results contribute to subsequent research on mosquito-virus interactions.

Enhanced bone regeneration via endochondral ossification using Exendin-4-modified mesenchymal stem cells.

Nonunions and delayed unions pose significant challenges in orthopedic treatment, with current therapies often proving inadequate. Bone tissue engineering (BTE), particularly through endochondral ossification (ECO), emerges as a promising strategy for addressing critical bone defects. This study introduces mesenchymal stem cells overexpressing Exendin-4 (MSC-E4), designed to modulate bone remodeling via their autocrine and paracrine functions. We established a type I collagen (Col-I) sponge-based in vitro model that effectively recapitulates the ECO pathway. MSC-E4 demonstrated superior chondrogenic and hypertrophic differentiation and enhanced the ECO cell fate in single-cell sequencing analysis. Furthermore, MSC-E4 encapsulated in microscaffold, effectively facilitated bone regeneration in a rat calvarial defect model, underscoring its potential as a therapeutic agent for bone regeneration. Our findings advocate for MSC-E4 within a BTE framework as a novel and potent approach for treating significant bone defects, leveraging the intrinsic ECO process.

Multidimensional characteristics of musculoskeletal pain and risk of hip fractures among elderly adults: the first longitudinal evidence from CHARLS.

BACKGROUND: Hip fractures are a major public health concern among middle-aged and older adults. It is important to understand the associated risk factors to inform health policies and develop better prevention strategies. Musculoskeletal pain is a possible implicating factor, being associated with physical inactivity and risk of falls. However, the association between musculoskeletal pain and hip fractures has not been clearly investigated. METHODS: A nationally representative sample of the Chinese population was obtained from the China Health and Retirement Longitudinal Study (CHARLS). The study collected patient information on their demographic characteristics, socioeconomic status, other health-related behavior, and history of musculoskeletal pain and hip fractures. Univariate and multivariate analyses were conducted to investigate the factors influencing the risk of hip fracture, including factors related to the individual and to musculoskeletal pain. P for trend test was performed to assess the trend of each continuous variable. The robustness and bias were assessed using the bootstrap method. Restricted cubic spline regression was utilized to identify linear or non-linear relationships. RESULTS: Among the 18,813 respondents, a total of 215 individuals reported that they have experienced a hip fracture. An increased risk of hip fracture was associated with the presence of waist pain and leg pain (P < 0.05), as well as with an increased number of musculoskeletal pain sites (P < 0.05). For individuals aged 65 and above, a significant association was found between age and the risk of hip fracture (P < 0.05). Furthermore, respondents with lower education level had a higher risk of hip fracture compared to those with higher education levels (P < 0.05). CONCLUSION: In the Chinese population, the risk of hip fracture was found to be associated with both the location and extent of musculoskeletal pain, as well as with other factors such as age and demographic characteristics. The findings of this study may be useful for informing policy development and treatment strategies, and provide evidence for comparison with data from other demographic populations.

Microplastics affect mosquito from aquatic to terrestrial lifestyles and are transferred to mammals through mosquito bites.

Microplastics (MPs) transfer from the environment to living organisms is a nonignorable global problem. As a complete metamorphosis insect, the larvae and adult Culex quinquefasciatus mosquito live in aquatic and terrestrial environments, respectively, where they easily access MPs. However, little is known about mosquitoes' potential role in MPs accumulation throughout ecosystems. Therefore, we conducted a study with different MPs particle sizes (0.1/1/10 µm) and concentrations (0.5/5/50 µg/mL) on Cx. quinquefasciatus to address this issue. Once exposed at the young larval stage, MPs could accompany the mosquitoes their entire life. The fluorescence signals of MPs in the larvae were mainly located in the intestines. Its intensity increased (from 3.72 × 106 AU to 5.45 × 107 AU) as the concentrations of MPs increases. The fluorescence signals of MPs were also detected in the blood and skin tissues of mice bitten by adult mosquitoes with MPs containing in their bodies. Mosquitos exposed to MPs showed longer larval pupation and eclosion time as well as lower adult body weight. In addition, MPs significantly reduced the lethal effect of pyrethroid insecticides (97.77 % vs. 48.88 %, p < 0.05) with 15.1 % removal of the deltamethrin concentration. After MPs exposure, the relative abundance of the Cx. quinquefasciatus gut microbiome, such as Wolbachia spp., Elizabethkingia spp., and Asaia spp., changed as the MPs size and concentration changes. Mosquitoes provide a new pathway for MPs accumulation and transfer to higher-level living organisms. Moreover, MPs significantly reduce the control effect of deltamethrin, providing new guidelines for mosquito insecticide application in MPs contamination circumstances.

Genome-wide identification of the B3 transcription factor family in pepper (Capsicum annuum) and expression patterns during fruit ripening.

In plants, B3 transcription factors play important roles in a variety of aspects of their growth and development. While the B3 transcription factor has been extensively identified and studied in numerous species, there is limited knowledge regarding its B3 superfamily in pepper. Through the utilization of genome-wide sequence analysis, we identified a total of 106 B3 genes from pepper (Capsicum annuum), they are categorized into four subfamilies: RAV, ARF, LAV, and REM. Chromosome distribution, genetic structure, motif, and cis-acting element of the pepper B3 protein were analyzed. Conserved gene structure and motifs outside the B3 domain provided strong evidence for phylogenetic relationships, allowing potential functions to be deduced by comparison with homologous genes from Arabidopsis. According to the high-throughput transcriptome sequencing analysis, expression patterns differ during different phases of fruit development in the majority of the 106 B3 pepper genes. By using qRT-PCR analysis, similar expression patterns in fruits from various time periods were discovered. In addition, further analysis of the CaRAV4 gene showed that its expression level decreased with fruit ripening and located in the nucleus. B3 transcription factors have been genome-wide characterized in a variety of crops, but the present study is the first genome-wide analysis of the B3 superfamily in pepper. More importantly, although B3 transcription factors play key regulatory roles in fruit development, it is uncertain whether B3 transcription factors are involved in the regulation of the fruit development and ripening process in pepper and their specific regulatory mechanisms because the molecular mechanisms of the process have not been fully explained. The results of the study provide a foundation and new insights into the potential regulatory functions and molecular mechanisms of B3 genes in the development and ripening process of pepper fruits, and provide a solid theoretical foundation for the enhancement of the quality of peppers and their selection and breeding of high-yield varieties.

Using A Google Web Search Analysis to Assess the Utility of ChatGPT in Stem Cell Therapy.

OBJECTIVE: Since its introduction, the use of ChatGPT has increased significantly for medically related purposes. However, current research has not captured its applications in providing information on stem cell therapy. To address this gap, the present study compared the effectiveness of ChatGPT to Google in answering medical questions related to stem cell therapy. METHODS: The search term "stem cell therapy" was used to perform a Google web search, and the top 20 frequently asked questions along with answers were recorded together with relevant website sources. Of these questions, the top 10 questions were separately entered into ChatGPT, and the answers and the sources were recorded. Then, the following statement was entered into ChatGPT: "Do a Google search with the search term 'stem cell therapy' and record 20 common questions related to the search term." After obtaining these questions, each question was separately entered into ChatGPT for an answer and source. RESULTS: A majority of the top 20 questions provided by Google were related to fact, whereas a majority of the questions provided by ChatGPT were related to policy. The answer sources used by Google were mostly drawn from medical practice, while those used by ChatGPT were mostly drawn from academic information. CONCLUSION: Compared to Google, ChatGPT exhibits stronger capabilities in promoting awareness of stem cell therapy. ChatGPT has the ability to eliminate misleading information by providing accurate and reliable answers. However, the responses provided by ChatGPT are still general in nature and cannot substitute academic sources for providing specialized knowledge.

Population genetics of Aedes albopictus in the port cities of Hainan Island and Leizhou Peninsula, China.

BACKGROUND: Aedes albopictus is an important vector of arboviral diseases, transmitting yellow fever, dengue fever, chikungunya and Zika. Monitoring its population genetic diversity and genetic differentiation has become essential for the control of infectious disease epidemics, especially in the functional areas of ports of entry. Population genetic monitoring of Ae. albopictus in the port area can help in the monitoring of port mosquito invasions and establishing port sanitary and quarantine measures to prevent the introduction and transmission of vector-borne diseases. METHODS: Seventeen populations of Ae. albopictus were collected from five port cities on Hainan Island and the Leizhou Peninsula, 8 populations were collected from port areas, 4 from urban areas and 5 from rural areas. Nine microsatellite loci and the mitochondrial COI gene were used to study the population genetic diversity, population genetic structure and interpopulation gene flow of Ae. albopictus. RESULTS: The nine microsatellite loci used were highly polymorphic, with an average PIC value of 0.768. The UPGMA genetic tree, STRUCTURE barplot and PCoA analyses showed that the 17 Ae. albopictus populations could be divided into three genetic groups. All 17 populations showed high haplotype diversity (Hd = 0.8069-0.9678) and formed 133 distinct haplotypes. These haplotypes can be divided into four genetic clades, but they are not associated with the geographical distribution of Ae. albopictus. Fst and Nm showed strong gene flow and little differentiation among populations. CONCLUSION: Ae. albopictus in port areas are not significantly different from urban and rural populations due to strong gene flow, which prevents differentiation and increases the genetic diversity of the populations. High genetic diversity facilitates mosquito adaptation to complex environmental changes, which is a challenge for vector-borne disease control in port areas.

SPINT2 is involved in the proliferation, migration and phenotypic switching of aortic smooth muscle cells: Implications for the pathogenesis of thoracic aortic dissection.

Thoracic aortic dissection (TAD) is a severe and extremely dangerous cardiovascular disease. Proliferation, migration and phenotypic switching of vascular smooth muscle cells (SMCs) are major pathogenetic mechanisms involved in the development of TAD. The present study was designed to investigate the expression and potential function of serine peptidase inhibitor Kunitz type 2 (SPINT2) in TAD. The gene expression profile data for ascending aorta from patients with TAD were downloaded from the GEO database with the accession number GSE52093. Bioinformatics analysis using GEO2R indicated that the differentially expressed SPINT2 was prominently decreased in TAD. The expression levels of SPINT2 mRNA and protein in aortic dissection specimens and normal aorta tissues were measured using reverse transcription-quantitative PCR and western blotting. SPINT2 expression was downregulated in clinical samples from aortic dissection specimens of patients with TAD compared with the corresponding expression noted in tissues derived from patients without TAD. In vitro, platelet-derived growth factor BB (PDGF-BB) was applied to induce the isolated primary mouse aortic SMC phenotypic modulation (a significant upregulation in the expression levels of synthetic markers), and the SMCs were infected with the adenoviral vector, Ad-SPINT2, to construct SPINT2-overexpressed cell lines. SMC viability was detected by an MTT assay and SMC proliferation was detected via the presence of Ki-67-positive cells (immunofluorescence staining). To explore the effects of SPINT2 on SMC migration, a wound healing assay was conducted. ELISA and western blotting assays were used to measure the content and expression levels of MMP-2 and MMP-9. The expression levels of vimentin, collagen I, α-SMA and SM22α were measured using western blotting. The PDGF-BB-induced proliferation and migration of SMCs were recovered by SPINT2 overexpression. The increase in the expression levels of SPINT2 reduced the expression levels of active matrix metalloproteinases (MMPs), MMP-2 and MMP-9. Overexpression of SPINT2 suppressed SMC switching from a contractile to a synthetic type, as evidenced by decreased vimentin and collagen I expression levels along with increased α-smooth muscle actin and smooth muscle protein 22-α expression levels. Furthermore, activation of ERK was inhibited in SPINT2-overexpressing SMCs. A specific ERK agonist, 12-O-tetradecanoylphorbol-13-acetate, reversed the SPINT2-mediated inhibition of SMC migration and the phenotypic switching. Collectively, the data indicated that SPINT2 was implicated in the proliferation, migration and phenotypic switching of aortic SMCs, suggesting that it may be involved in TAD progression.

Cell fusing agent virus isolated from Aag2 cells does not vertically transmit in Aedes aegypti via artificial infection.

BACKGROUND: Cell fusing agent virus (CFAV) was the first insect-specific virus to be characterized, and has been reported to negatively influence the growth of arboviruses such as dengue, Zika, and La Cross, making it a promising biocontrol agent for mosquito-borne disease prevention. Aedes aegypti Aag2 cells were naturally infected with CFAV. However, the ability of this virus to stably colonize an Ae. aegypti population via artificial infection and how it influences the vector competence of this mosquito have yet to be demonstrated. METHODS: CFAV used in this study was harvested from Aag2 cells and its complete genome sequence was obtained by polymerase chain reaction and rapid amplification of complementary DNA ends, followed by Sanger sequencing. Phylogenetic analysis of newly identified CFAV sequences and other sequences retrieved from GenBank was performed. CFAV stock was inoculated into Ae. aegypti by intrathoracic injection, the survival of parental mosquitoes was monitored and CFAV copies in the whole bodies, ovaries, and carcasses of the injected F0 generation and in the whole bodies of the F1 generation on different days were examined by reverse transcription-quantitative polymerase chain reaction. RESULTS: The virus harvested from Aag2 cells comprised a mixture of three CFAV strains. All genome sequences of CFAV derived from Aag2 cells clustered into one clade but were far from those isolated or identified from Ae. aegypti. Aag2-derived CFAV efficiently replicated in the mosquito body and did not attenuate the survival of Ae. aegypti. However, the viral load in the ovarian tissues was much lower than that in other tissues and the virus could not passage to the offspring by vertical transmission. CONCLUSIONS: The results of this study demonstrate that Aag2-derived CFAV was not vertically transmitted in Ae. aegypti and provide valuable information on the colonization of mosquitoes by this virus.

Evolution of knockdown resistance (kdr) mutations of Aedes aegypti and Aedes albopictus in Hainan Island and Leizhou Peninsula, China.

Background: Aedes aegypti and Aedes albopictus are important vectors of human arboviruses, transmitting arboviral diseases such as yellow fever, dengue, chikungunya and Zika. These two mosquitoes coexist on Hainan Island and the Leizhou Peninsula in China. Over the past 40 years, the distribution of Ae. albopictus has gradually expanded in these areas, while the distribution of Ae. aegypti has declined dramatically mainly due to the ecological changes and some other factors such as heavy use of insecticide indoor based on endophagic bloodfeeding of the species. Methods: This study focused on the knockdown resistance (kdr) genes of both mosquitoes, investigated their mutations, and analyzed their haplotype and evolutionary diversity combined with population genetic features based on the ND4/ND5 genes to further elucidate the molecular mechanisms underlying the development of insecticide resistance in both mosquitoes. Results: Three mutations, S989P, V1016G and F1534C, were found to be present in Ae. aegypti populations, and the three mutations occurred synergistically. Multiple mutation types (F1534C/S/L/W) of the F1534 locus are found in Ae. albopictus populations, with the three common mutations F1534C, F1534S and F1534L all having multiple independent origins. The F1534W (TTC/TGG) mutation is thought to have evolved from the F1534L (TTC/TTG) mutation. The F1534S (TTC/TCG) mutation has evolved from the F1534S (TTC/TCC) mutation. The most common form of mutation at the F1534 locus found in this study was S1534C, accounting for 20.97%, which may have evolved from the F1534C mutation. In addition, a new non-synonymous mutation M1524I and 28 synonymous mutations were identified in Ae. albopictus populations. Correlation analysis showed that the genetic diversity of Ae. aegypti and Ae. albopictus populations did not correlate with their kdr haplotype diversity (P>0.05), but strong gene flow between populations may have contributed to the evolution of the kdr gene. Conclusion: The study of kdr gene evolution in the two mosquito species may help to identify the evolutionary trend of insecticide resistance at an early stage and provide a theoretical basis for improving the efficiency of biological vector control and subsequent research into new insecticides.

Genetic diversity of Aedes aegypti and Aedes albopictus from cohabiting fields in Hainan Island and the Leizhou Peninsula, China.

BACKGROUND: Aedes aegypti and Ae. albopictus are important human arbovirus vectors that can spread arboviral diseases such as yellow fever, dengue, chikungunya and Zika. These two mosquito species coexist on Hainan Island and the Leizhou Peninsula in China. Over the past 40 years, the distribution of Ae. albopictus in these areas has gradually expanded, while Ae. aegypti has declined sharply. Monitoring their genetic diversity and diffusion could help to explain the genetic influence behind this phenomenon and became key to controlling the epidemic of arboviruses. METHODS: To better understand the genetic diversity and differentiation of these two mosquitoes, the possible cohabiting areas on Hainan Island and the Leizhou Peninsula were searched between July and October 2021, and five populations were collected. Respectively nine and 11 microsatellite loci were used for population genetic analysis of Ae. aegypti and Ae. albopictus. In addition, the mitochondrial coxI gene was also selected for analysis of both mosquito species. RESULTS: The results showed that the mean diversity index (PIC and SI values) of Ae. albopictus (mean PIC = 0.754 and SI = 1.698) was higher than that of Ae. aegypti (mean PIC = 0.624 and SI = 1.264). The same results were also observed for the coxI gene: the genetic diversity of all populations of Ae. albopictus was higher than that of Ae. aegypti (total H = 45 and Hd = 0.89958 vs. total H = 23 and Hd = 0.76495, respectively). UPGMA dendrogram, DAPC and STRUCTURE analyses showed that Ae. aegypti populations were divided into three clusters and Ae. albopictus populations into two. The Mantel test indicated a significant positive correlation between genetic distance and geographic distance for the Ae. aegypti populations (R2 = 0.0611, P = 0.001), but the correlation was not significant for Ae. albopictus populations (R2 = 0.0011, P = 0.250). CONCLUSIONS: The population genetic diversity of Ae. albopictus in Hainan Island and the Leizhou Peninsula was higher than that of Ae. aegypti. In terms of future vector control, the most important and effective measure was to control the spread of Ae. albopictus and monitor the population genetic dynamics of Ae. aegypti on Hainan Island and the Leizhou Peninsula, which could theoretically support the further elimination of Ae. aegypti in China.

Population genetic characterization of (Aedes albopictus) mosquitoes (Diptera: Culicidae) from the Yangtze River Basin of China based on rDNA-ITS2.

BACKGROUND: Aedes albopictus is an important vector of many mosquito-borne viral diseases, including dengue fever and Zika. In recent years, it has spread and colonized tropical, subtropical and temperate regions worldwide. Monitoring of Ae. albopictus population dynamics is an important tool for early warning of mosquito-borne infections. Because the genetic diversity and genetic structure of natural populations are the genetic bases of population dynamics, studies of population genetics can reveal the origin, differentiation and dispersal characteristics of Ae. albopictus populations. Then, their evolutionary potential and environmental adaptability can be analyzed, providing a theoretical basis for the formulation of accurate Ae. albopictus surveillance and integrated control programs. METHODS: In 2018, 552 Ae. albopictus larvae were collected during an invasive mosquito species surveillance project in China's Yangtze River Basin. Morphological analysis was performed to assign the adult mosquitoes to species, and then the genetic marker ITS2 was amplified and sequenced. RESULTS: There were 179 haplotypes among 552 ITS2 sequences. In total, 155/179 (86.59%) haplotypes were specific to individual populations, and 24/179 (13.41%) haplotypes were shared by populations. Hap4 (126), Hap7 (43), and Hap16 (34) were the most numerous haplotypes and the most widely distributed. The overall Hd was 0.928, π was 0.031, the mean nucleotide difference number (K) was 7.255, and the number of segregating sites was 169. TCS network maps mainly showed a single star-like scattered distribution. According to geographical location, there were no obvious haplotype groups, and the haplotypes were intricately connected. The genetic diversity of Ae. albopictus populations in the Yangtze River Basin was high. The molecular variance observed in the populations of Ae. albopictus mainly occurred among individuals within populations, accounting for 98.79% of the total, while that among populations accounted for only 1.21% of the total. Only the populations of Ae. albopictus in the Chongqing and Sichuan regions showed a moderate degree of population genetic differentiation, while genetic differentiation between the other regions were small, gene exchange was very common, and genetic differentiation within populations was minimal. CONCLUSIONS: According to this study, the genetic diversity of Ae. albopictus populations in the Yangtze River Basin is high, the genetic differentiation among populations is small, and gene exchange is common. In addition, frequent interregional exchange exacerbates the abnormal spread of vectors. This study highlighted the potential spread route of the vector Ae. albopictus in the Yangtze River Basin. There are three potential dispersal routes for Ae. albopictus populations in the Yangtze River Basin. The findings could be helpful for effective surveillance and early warning of Ae. albopictus vectors.

The predicted potential distribution of Aedes albopictus in China under the shared socioeconomic pathway (SSP)1-2.6.

Aedes albopictus (Skuse) (Diptera: Culicidae) is one of the 100 most invasive species in the world and represents a significant threat to public health. The distribution of Ae. albopictus has been expanding rapidly due to increased international trade, population movement, global warming and accelerated urbanization. Consequently, it is very important to know the potential distribution area of Ae. albopictus in advance for early warning and control of its spread and invasion. We randomly selected 282 distribution sites from 27 provincial-level administrative regions in China, and used the GARP and MaxEnt models to analyze and predict the current and future distribution areas of Ae. albopictus in China. The results showed that the current range of Ae. albopictus in China covers most provinces such as Yunnan and Guizhou Provinces, and the distribution of Ae. albopictus in border provinces such as Tibet, Gansu and Jilin Provinces tend to expand westwards. In addition, the potential distribution area of Ae. albopictus in China will continue to expand westwards due to future climate change under the SSP126 climate scenario. Furthermore, the results of environmental factor filtering showed that temperature and precipitation had a large effect on the distribution probability of Ae. albopictus.