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Background: Numerous approaches have been utilized to optimize mesenchymal stem cells (MSCs) performance in treating osteoarthritis (OA), however, the constrained diminished activity and chondrogenic differentiation capacity impede their therapeutic efficacy. Previous investigations have successfully shown that pretreatment with nanosecond pulsed electric fields (nsPEFs) significantly enhances the chondrogenic differentiation of MSCs. Therefore, this study aims to explore nsPEFs as a strategy to improve OA therapy by enhancing MSCs' activity and chondrogenic differentiation and also investigate its potential mechanism. Methods: In this study, a million MSCs were carefully suspended within a 0.4-cm gap cuvette and subjected to five pulses of nsPEFs (100 ns at 10 kV/cm, 1 Hz), with a 1-s interval between each pulse. A control group of MSCs was maintained without nsPEFs treatment for comparative analysis. nsPEFs were applied to regulate the MSCs performance and hinder OA progresses. In order to further explore the corresponding mechanism, we examined the changes of MSCs transcriptome after nsPEF pretreatment. Finally, we studied the properties of extracellular vesicles (EVs) secreted by MSCs affected by nsPEF and the therapeutic effect on OA. Results: We found that nsPEFs pretreatment promoted MSCs migration and viability, particularly enhancing their viability temporarily in vivo, which is also confirmed by mRNA sequencing analysis. It also significantly inhibited the development of OA-like chondrocytes in vitro and prevented OA progression in rat models. Additionally, we discovered that nsPEFs pretreatment reprogrammed MSC performance by enhancing EVs production (5.77 ± 0.92 folds), and consequently optimizing their therapeutic potential. Conclusions: In conclusion, nsPEFs pretreatment provides a simple and effective strategy for improving the MSCs performance and the therapeutic effects of MSCs for OA. EVs-nsPEFs may serve as a potent therapeutic material for OA and hold promise for future clinical applications. The translational potential of this article: This study indicates that MSCs pretreated by nsPEFs greatly inhibited the development of OA. nsPEFs pretreatment will be a promising and effective method to optimize the therapeutic effect of MSCs in the future.

Background: Platelet-rich plasma (PRP) helps promote wound healing, but it is unclear whether it stimulates breast cancer cell proliferation, which restricts its application in breast cancer patients. This article explored the effect of PRP on breast cancer cell proliferation through preclinical experiments. Method: We cultivated MDA-MB-231 breast cancer cells with PRP in vitro. Subsequently, we employed Cell Counting Kit-8 (CCK-8) assays to assess their proliferation ability, wound healing assays to evaluate their migration ability, and Transwell assays to detect their invasion ability. Mouse breast cancer 4T1 cells were subcutaneously inoculated into nude mice. After tumor formation, PRP was injected around each tumor. Tumor size was measured. After 15 days, the tumors were surgically removed. Immunohistochemistry was used to detect key proteins involved in proliferation and apoptosis. Results: PRP inhibited the proliferation, migration, and invasion of MDA-MB-231 in vitro. After PRP was injected around tumors in nude mice, tumor growth slowed, Ki67 and phospho-histone H3 (pHH3) expression decreased, and Caspase 3 and Poly (adenosine diphosphate-ribose) polymerase 1 (PARP1) expression increased. Conclusion: The PRP inhibits the proliferation of breast cancer MDA-MB-231 and 4T1 cells.

Aedes albopictus, a common mosquito in Zhejiang Province, is a carrier of more than twenty arboviruses. There are dozens or even hundreds of imported cases of dengue fever every year in Zhejiang Province, and there have also been many local outbreaks caused by imported cases of dengue fever. The objectives were to assess the resistance of larvae and adults of several Ae. albopictus strains in Zhejiang Province to commonly used pyrethroid insecticides (beta-cypermethrin, deltamethrin and permethrin), and detect mutations in the sodium channel gene, to further analyse the relationship between phenotypic resistance and the frequency of mutations. The resistance of eight field strains of Ae. albopictus larvae to beta-cypermethrin, deltamethrin and permethrin ranged from 8.17 to 36.06, 12.12–107.3 and 1.55–81.9, respectively, and there was a significant positive correlation of interaction resistance among the three insecticides. The mutation frequencies of I1532T and F1534S in the larvae of Ae. albopictus were 0–6.25 % and 42.19–100.00 %. Moreover, the diagnostic doses of the three pyrethroids for adult Ae. albopictus mosquitoes were 0.2510 g/L, 0.1562 g/L, and 0.9072 g/L. Except for the Zhoushan strain, which was suspected to be resistant to beta-cypermethrin, the other field strains were resistant to the three pyrethroids, and there was a significant positive correlation of cross-resistance among the three insecticides. The mutation frequencies of I1532T and F1534S of adult Ae. albopictus were 0–1.56 % and 62.50–100.00 %. In addition, the LC50 of the larvae and the mortality rate of adult Ae. albopictus after treatment with the three pyrethroids were significantly and positively correlated with the frequency of the F1534S mutation. F1534S mutation occurred earlier than I1532T mutation in both larvae and adult Ae. albopictus. F1534S mutation in the sodium channel gene may be a particular biomolecular detection marker for resistance to pyrethroid insecticides in Ae. albopictus in Zhejiang Province.

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.

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.

Summary: Many countries and commercial organizations have shown great interest in constructing a Martian base. In situ resource utilization (ISRU) provides a cost-effective way to achieve this ambitious goal. In this article, we proposed to use Martian soil simulant to produce a fiber to satisfy material requirement for the construction of Martian base. The composition, melting behavior, and fiber forming process of the soil simulant was studied, and continuous fiber with maximum strength of 1320 MPa and elastic modulus of 99 GPa was obtained on a spinning facility. The findings of this study demonstrate the feasibility of ISRU to prepare Martian fiber from the soil on the Mars, offering a new way to obtain key materials for the construction of a Martian base.

Abstract 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

Abstract 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.

BackgroundTo 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.MethodsThis 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.ResultsUsing 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.ConclusionThis 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.

Abstract 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. Graphical abstract

Stem cells are widely used in scientific research because of their ability to self-renew and differentiate into a variety of specialized cell types needed for body functions. However, the self-renewal and differentiation of stem cells are regulated by various stimuli, with mechanical stimulation being particularly notable due to its ability to mimic the physical environment in the body. This study systematically collected 2638 research papers published between 1994 and 2024, employing tools such as VOSviewer, CiteSpace, and GraphPad Prism to uncover research hotspots, publication trends, and collaboration networks. The results indicate a yearly increase in global research on the shear stress stimulation of stem cells, with significant contributions from the United States and China in terms of research investment and output. Future research directions include a deeper understanding of the mechanisms underlying mechanical stimulation’s effects on stem cell differentiation, the development of new materials and scaffold designs to better replicate the natural cellular environment, and advancements in regenerative medicine. Despite considerable progress, challenges remain in translating basic research findings into clinical applications.

Abstract 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 (R 2 = 0.0611, P = 0.001), but the correlation was not significant for Ae. albopictus populations (R 2 = 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. Graphical Abstract

The regeneration of hierarchical osteochondral units is challenging due to difficulties in inducing spatial, directional and controllable differentiation of mesenchymal stem cells (MSCs) into cartilage and bone compartments. Emerging organoid technology offers new opportunities for osteochondral regeneration. In this study, we developed gelatin-based microcryogels customized using hyaluronic acid (HA) and hydroxyapatite (HYP), respectively for inducing cartilage and bone regeneration (denoted as CH-Microcryogels and OS-Microcryogels) through in vivo self-assembly into osteochondral organoids. The customized microcryogels showed good cytocompatibility and induced chondrogenic and osteogenic differentiation of MSCs, while also demonstrating the ability to self-assemble into osteochondral organoids with no delamination in the biphasic cartilage-bone structure. Analysis by mRNA-seq showed that CH-Microcryogels promoted chondrogenic differentiation and inhibited inflammation, while OS-Microcryogels facilitated osteogenic differentiation and suppressed the immune response, by regulating specific signaling pathways. Finally, the in vivo engraftment of pre-differentiated customized microcryogels into canine osteochondral defects resulted in the spontaneous assembly of an osteochondral unit, inducing simultaneous regeneration of both articular cartilage and subchondral bone. In conclusion, this novel approach for generating self-assembling osteochondral organoids utilizing tailor-made microcryogels presents a highly promising avenue for advancing the field of tissue engineering.

Aedes aegypti (Ae. aegypti) is a major vector of dengue virus (DENV) and Zika virus (ZIKV). Understanding the complex interaction mechanisms between mosquito vectors and arboviruses is essential to interrupt virus transmission. This study constructed CYP4C21 knockout (KO) Aag2 cells (Ae. aegypti cells) and confirmed that CYP4C21 KO reduced DENV2 and ZIKV copies in Aag2 cells, which suggests that CYP4C21 may play an important role in mosquito infection with arboviruses. Furthermore, it is the first report of the CYP4 family related to viral infection, which lays the foundation for exploring the role of the CYP4C21 in the interaction of Ae. aegypti and arbovirus and provides novel insights into the function of cytochrome family proteins.

Abstract Background Musculoskeletal pain is a major cause of physical disability, associated with huge socioeconomic burden. Patient preference for treatment is an important factor contributing to the choice of treatment strategies. However, effective measurements for evaluating the ongoing management of musculoskeletal pain are lacking. To help improve clinical decision making, it’s important to estimate the current state of musculoskeletal pain management and analyze the contribution of patient treatment preference. Methods A nationally representative sample for the Chinese population was derived from the China Health and Retirement Longitudinal Study (CHARLS). Information on the patients’ demographic characteristics, socioeconomic status, other health-related behavior, as well as history on musculoskeletal pain and treatment data were obtained. The data was used to estimate the status of musculoskeletal pain treatment in China in the year 2018. Univariate analysis and multivariate analysis were used to find the effect factors of treatment preference. XGBoost model and Shapley Additive exPlanations (SHAP) method were performed to analyze the contribution of each variable to different treatment preferences. Results Among 18,814 respondents, 10,346 respondents suffered from musculoskeletal pain. Approximately 50% of musculoskeletal pain patients preferred modern medicine, while about 20% chose traditional Chinese medicine and another 15% chose acupuncture or massage therapy. Differing preferences for musculoskeletal pain treatment was related to the respondents’ gender, age, place of residence, education level, insurance status, and health-related behavior such as smoking and drinking. Compared with upper or lower limb pain, neck pain and lower back pain were more likely to make respondents choose massage therapy (P

Background Neuroarthropathy of the knee or Charcot knee, leading to chronic joint destruction, is a rare disease that is difficult to diagnose. The treatment of this condition is difficult and controversial. Case Presentation A 74‐year‐old Asian woman has had bilateral knee pain for 22 years and deformity for 10 years, which has been aggravating for 2 months. Physical examination showed bilateral knee varus deformity greater than 15°, and −20 to 90° range of motion. X‐ray revealed bilateral varus deformity with massive free body hyperplasia. Combined with medical history as syringomyelia, the patient was diagnosed with bilateral Charcot knees and bilateral joint replacements were performed using Legacy Constrained Condylar Knee prostheses (LCCK; Zimmer, USA). The patient reported satisfactory treatment outcomes, pain relief, and improved range of motion in both knees, without postoperative complications or prosthesis loosening at 2 year after operation. Conclusions Total knee arthroplasty (TKA) may be considered a viable option for treating the Charcot knee. The use of constrained condylar prostheses can produce satisfactory results. Attention should be given to survival risks, complications, and other potential determining factors associated with TKA when devising a treatment strategy for the Charcot knee.

Objective Prosthetic joint infection (PJI) is the main reason of failure of total joint arthroplasty (TJA). This study aimed to investigate the global trends and network visualization in research of PJI. Methods Publications in PJI search during 1980–2022 were extracted from the Science Citation Index‐Expanded of Web of Science Core Collection database (WoSCC). The source data was investigated and analyzed by bibliometric methodology. For network visualization, VOS viewer and R software was used to perform bibliographic coupling, co‐citation, co‐authorship and co‐occurrence analysis and to predict the publication trends in PJI research. Results There were 7288 articles included. The number of publications and relative research interests increased gradually per year globally. The USA made the highest contributions in the world and with the highest H‐index and the most citations. Journal of Arthroplasty published the highest number of articles in this area. The Mayo Clinic, Thomas Jefferson University (Rothman Institute), Hospital Special Surgery and the Rush University were the most contributive institutions by network visualization. Included studies were divided into four clusters: bacterial pathogenic mechanism and antibacterial drugs study, TJA complications, risk factors and epidemiology of PJI, diagnosis of PJI, and revision surgical management. More articles in PJI could be published over the next few years. Conclusion The number of publications about PJI will be increasing dramatically based on the global trends and network visualization. The USA made the highest contributions in PJI. Diagnosis and revision management may be the next hot spots in this field.

Objective Musculoskeletal pain is the most prominent clinical manifestation of more than 150 musculoskeletal disease conditions, and its effective long‐term management poses a great challenge to healthcare systems globally. For this, it is important to understand current research progress on musculoskeletal pain management. The purpose of the present study is to provide a comprehensive insight into the current state of research and global trends in musculoskeletal pain management. Methods Publications on musculoskeletal pain management from 1972 to 2021 were retrieved from the Science Citation Index‐Expanded (SCIE) database. Included articles were any article type related to aspects of musculoskeletal pain management, including etiology, mechanisms, epidemiology, treatment, outcomes, side effects, and patient compliance. Publication data were analyzed using bibliometric methods. The software VOSviewer was employed to perform bibliographic coupling, co‐authorship, co‐citation, and co‐occurrence analysis, and to visualize publication tendencies in musculoskeletal pain management. Results A total of 5475 articles were included in this study. The number of global publications on musculoskeletal pain management has escalated annually. Based on the number of publications and citations from the published literature, as well as the H‐index, the United States led global contributions in this area. The institutions making the highest contributions were the League of European Research Universities (LERU), the University of Sydney, and Harvard University. The journal BMC Musculoskeletal Disorders published the highest number of articles in this area. The published studies fall under six groups: “Prevention and rehabilitation,” “Etiology and diagnosis,” “Clinical study,” “Epidemiology,” “Mental health,” and “Education.” High‐quality primary studies and epidemiology are predicted to be the next prevailing topics in this field of research. Conclusions Based on current global trends, the number of publications on musculoskeletal pain management will continue to increase. Future studies will likely place more emphasis on high‐quality randomized controlled trials (RCTs) and epidemiological studies.

Abstract Mosquito transmit numbers of parasites and pathogens resulting in fatal diseases. Species identification is a prerequisite for effective mosquito control. Existing morphological and molecular classification methods have evitable disadvantages. Here we introduced Deep learning techniques for mosquito species identification. A balanced, high-definition mosquito dataset with 9900 original images covering 17 species was constructed. After three rounds of screening and adjustment-testing (first round among 3 convolutional neural networks and 3 Transformer models, second round among 3 Swin Transformer variants, and third round between 2 images sizes), we proposed the first Swin Transformer-based mosquito species identification model (Swin MSI) with 99.04% accuracy and 99.16% F1-score. By visualizing the identification process, the morphological keys used in Swin MSI were similar but not the same as those used by humans. Swin MSI realized 100% subspecies-level identification in Culex pipiens Complex and 96.26% accuracy for novel species categorization. It presents a promising approach for mosquito identification and mosquito borne diseases control.

Objective Musculoskeletal pain is having growing impacts worldwide with clinical challenge in pain management. The purpose of the present study is to investigate the preferences of orthopedic surgeons of China for using medicine in musculoskeletal pain. Methods A questionnaire was developed, including the following domains, personal information, medication preference for pain treatment, and perceptions of topical medicine. Ten participants were selected to confirm the consistency of questionnaire. A cross‐sectional survey was conducted in orthopedic physicians with different specialties in different regions of China via the online survey platform. The participants' survey results were analyzed one‐way and multi‐way using chi‐square test and logistic regression. Results The pre‐survey analysis results of 10 randomly selected investigators were a mean weighted kappa coefficient of 0.76 (range 0.61–0.89), which indicated the substantial consistency of the present questionnaire. A total of 1099 orthopedic surgeons (mean age, 41.67 ± 8.31 years) responded to our survey, most of whom were male (90.72%), and most of whom worked in level III hospitals (63.24%) and trained in modern medicine (71.43%). Most surgeons who participated in the survey had used topical analgesics in their clinical work (95.81%), and most preferred to use topical analgesics (39.50%) or a combination of oral analgesics (28.87%). Primary reasons for preferring topical analgesics were as follows: less adverse reactions (68.01%); ease of use (60.90%); and not interfering with other oral medications (49.60%). The preference for prescribing topical analgesics increased with the education level of the respondent, where statistically significant differences were seen (P

Abstract Background Both Culex quinquefasciatus and Cx. pipiens molestus are sibling species within Cx. pipiens complex. Even though they are hard to distinguish morphologically, they have different physiological behaviors. However, the molecular mechanisms underlying these differences remain poorly understood. Methods Transcriptome sequencing was conducted on antennae of two sibling species. The identification of the differentially expressed genes (DEGs) was performed by the software DESeq2. Database for Annotation, Visualization and Integrated Discovery was used to perform GO pathway enrichment analysis. The protein–protein interaction (PPI) network was constructed with Cytoscape software. The hub genes were screened by the CytoHubba plugin and Degree algorithms. The identified genes were verified by quantitative real-time PCR. Results Most annotated transcripts (14,687/16,005) were expressed in both sibling species. Among 15 identified odorant-related DEGs, OBP10 was expressed 17.17 fold higher in Cx. pipiens molestus than Cx. quinquefasciatus. Eighteen resistance-related DEGs were identified, including 15 from CYP gene family and three from acetylcholinesterase, in which CYP4d1 was 86.59 fold more highly expressed in C. quinquefasciatus. Three reproductive DEGs were indentified with the expression from 5.01 to 6.55 fold. Among eight vision-related DEGs, retinoic acid receptor RXR-gamma in Cx. pipiens molestus group was more expressed with 214.08 fold. Among the 30 hub genes, there are 10 olfactory-related DEGs, 16 resistance-related DEGs, and four vision-related DEGs, with the highest score hub genes being OBP lush (6041148), CYP4C21 (6044704), and Rdh12 (6043932). The RT-qPCR results were consistent with the transcriptomic data with the correlation coefficient R = 0.78. Conclusion The study provided clues that antennae might play special roles in reproduction, drug resistance, and vision, not only the traditional olfactory function. OBP lush, CYP4C21, and Rdh12 may be key hints to the potential molecular mechanisms behind the two sibling species' biological differences. Graphical Abstract

BackgroundAedes 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.MethodsThis 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.ResultsThree 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.ConclusionThe 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.

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.

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.

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.

Objective Predicting the successful preservation of posterior cruciate ligament (PCL) in total knee arthroplasty (TKA) is an important step for preoperative planning to secure the satisfactory outcomes. We aimed to examine the preoperative factors predicting the successful preservation of the PCL in cruciate‐retaining TKA and the outcome of sacrificing the PCL. Methods In this retrospective study, we analyzed TKAs consecutively performed by a single surgeon between January 2019 and August 2021 who had been preoperatively planned to undergo implantation of cruciate‐retaining (CR) prostheses. The outcome of the current study was whether the PCL was retained or sacrificed. Anterior‐stabilized (AS) tibial bearings when the PCL was sacrificed as needed were used intraoperatively. Age, sex, body mass index (BMI), and preoperative diagnosis from the patients' medical records were obtained. The medial‐lateral width of epicondyle (MLW), the medial posterior condyle height (MPCH), the lateral posterior condyle height (LPCH), the ratio of MLW and MPCH, the ratio of MLW and LPCH, the Insall–Salvati index, and the severity of the varus or valgus deformity were measured using preoperative radiographs. Univariate and multivariate regression were fitted to assess the association of these factors with the successful retention of PCL. To examine the influence of sacrifice of the PCL on the surgical procedure, the size of the tibial and femoral components, the thickness of the polyethylene insert, and the rate of patella replacement between the CR group and AS group were also compared using t tests or chi‐square tests. Results Among 307 TKAs included, PCL was sacrificed with concurrent use of AS prostheses in 89 (29.0%) procedures. Knees with rheumatoid arthritis (P

Background Periprosthetic osteolysis is a serious complication following total hip arthroplasty (THA). However, most orthopedic surgeons only focus on bone loss and hip reconstruction. Thus, it was required to understand the treatment algorithm for periprosthetic osteolysis integrally. Case Presentation A 52‐year‐old Asian male presented with chronic hip pain. A mass appeared on the medial side of the proximal left thigh at more than 20 years after bilateral THA. Radiographs revealed catastrophic periprosthetic osteolysis, especially on the acetabular side. Large amounts of necrotic tissue and bloody fluids were thoroughly debrided during revision THA. A modular hemipelvic prosthesis was used for revision of the left hip. Four years later, the patient presented with right hip pain, where a mass appeared on the medial side of the proximal right thigh. A primary acetabular implant with augment was used for revision of the right hip. Laboratory evaluation of bloody fluid retrieved from surgery revealed elevated levels of inflammatory markers. Conclusion Inflammatory responses to polyethylene wear debris can lead to severe bone resorption and aseptic loosening in the long‐term following THA. Therefore, in spite of revision THA, interrupting the cascade inflammatory might be the treatment principle for periprosthetic osteolysis.

The self-repair ability of cartilage defects is limited, and 3D printing technology provides hope for the repair and regeneration of cartilage defects. Although 3D printing technology and cartilage repair and regeneration have been studied for decades, there are still few articles specifically describing the relationship between 3D printing and cartilage defect repair and regeneration, and a bibliometric analysis has not been completed. To supplement, sort out and summarize the content in related fields, we analyzed the research status of 3D printing technology and cartilage repair and regeneration from 2002 to 2022. According to the set search strategy, the Web of Science Core Collection was used as the data source, and the literature search was completed on December 6, 2022. CiteSpace V and VOSviewer were used as bibliometric tools to complete the analysis of the research focus and direction of the published literature. Based on the analysis results, we focus on the occurrence and development of this field of combined medical and engineering research. Moreover, the current advantages and limitations of this field as well as future development prospects are discussed in depth. It will help to shape researchers’ understanding of 3D printing and cartilage repair and regeneration, inspire researchers’ research ideas, guide research directions, and promote related research results to clinical application.

Abstract Background Aedes aegypti is one of the most important vectors of zoonotic diseases worldwide, and its survival and reproductive processes depend heavily on its olfactory system. In this study, the expression levels of all odorant receptor (OR) genes of Ae. aegypti were explored during different physiological periods to identify olfactory genes that may be associated with mosquito blood-feeding and the search for oviposition sites. Methods Four experimental groups, consisting of Ae. aegypti males, pre-blood-feeding females, post-blood-feeding females and post-oviposition females, were established. A total of 114 pairs of primers targeting all messenger RNA encoded by OR genes were designed based on the whole genome of Ae. aegypti. The expression of OR genes was evaluated by real-time fluorescence quantitative PCR for relative quantification and the comparison of differences between groups. Results A total of 53 differentially expressed OR genes were identified between males and females in Ae. aegypti antennae. Also, eight, eight and 13 differentially expressed OR genes were identified in pre- versus post-blood-feeding females, in pre- versus post-oviposition females and in post-blood-feeding versus post-oviposition females, respectively. In addition, 16 OR genes were significantly differentially expressed in multiple physiological periods of the mosquitoes. Conclusions A large number of ORs with significant intergroup differences and high expression levels were screened in this study. Some of these genes are reported for the first time, providing possible targets for the development of mosquito control pathways based on the olfactory system. Graphical Abstract

Objective To describe a non‐anatomical arthroscopic all‐inside repair of medial meniscus posterior root tear (MMPRT) to posterior cruciate ligament (PCL) technique for patients with normal lower limb alignment and to evaluate the short‐term clinical and radiologic outcomes. Methods MMPRT directly to PCL was repaired with all‐inside horizontal mattress suturing technique rather than by the transtibial pullout suture technique or anchor suturing repair technique in 20 Laparade Type II MMPRT patients with normal lower limb alignment during 2018–2019. The clinical and radiological outcomes were evaluated retrospectively for at least 2 years follow‐up. The VAS score, Lysholm score, Tegner activity score were evaluated preoperatively and at the final follow‐up. The status of the medial meniscus posterior root were assessed on magnetic resonance imaging (MRI) preoperatively and at the final follow‐up. Results Twenty patients (mean age 54.5 ± 19.5 years) were included in the present study. The mean follow‐up duration was 32.5 ± 5.8 months. The VAS score was significantly decreased from preoperative 6.5 ± 1.5 to 2.1 ± 1.4 at the final follow‐up (P

Mesenchymal stem cells (MSCs) can be effective in alleviating the progression of osteoarthritis (OA). However, low MSC retention and survival at the injection site frequently require high doses of cells and/or repeated injections, which are not economically viable and create additional risks of complications. In this study, we produced MSC-laden microcarriers in spinner flask culture as cell delivery vehicles. These microcarriers containing a low initial dose of MSCs administered through a single injection in a rat anterior cruciate ligament (ACL) transection model of OA achieved similar reparative effects as repeated high doses of MSCs, as evaluated through imaging and histological analyses. Mechanistic investigations were conducted using a co-culture model involving human primary chondrocytes grown in monolayer, together with MSCs grown either within 3D constructs or as a monolayer. Co-culture supernatants subjected to secretome analysis showed significant decrease of inflammatory factors in the 3D group. RNA-seq of co-cultured MSCs and chondrocytes using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed processes relating to early chondrogenesis and increased extracellular matrix interactions in MSCs of the 3D group, as well as phenotypic maintenance in the co-cultured chondrocytes. The cell delivery platform we investigated may be effective in reducing the cell dose and injection frequency required for therapeutic applications.

Highlights 1. RNA sequencing showed that OBP28 gene expression was higher in the deltamethrin-resistant strains. 2. The qPCR results verified that OBP28 gene expression was higher in the resistant strains. 3. RNA interference silenced the OBP28 gene at an efficiency of more than 50%. 4. The sensitivity of Cx. quinquefasciatus to deltamethrin increased after OBP28 had been silenced.

Introduction: Since Aedes aegypti invaded Yunnan Province in 2002, its total population has continued to expand. Shi et al. used microsatellite and mitochondrial molecular markers to study the Ae. aegypti populations in Yunnan Province in 2015 and 2016, found that it showed high genetic diversity and genetic structure. However, there are few studies on the population genetic characteristics of Ae. aegypti in Yunnan Province under different levels of human intervention. This study mainly used two common types of molecular markers to analyze the genetic characteristics of Ae. aegypti, revealing the influence of different input, prevention and control pressures on the genetic diversity and structure of this species. Understanding the genetic characteristics of Ae. aegypti populations and clarifying the diversity, spread status, and source of invasion are essential for the prevention, control and elimination of this disease vector.Methods: We analyzed the genetic diversity and genetic structure of 22 populations sampled in Yunnan Province in 2019 and 17 populations sampled in 2020 through nine microsatellite loci and COI and ND4 fragments of mitochondrial DNA. In 2019, a total of 22 natural populations were obtained, each containing 30 samples, a total of 660 samples. In 2020, a total of 17 natural populations were obtained. Similarly, each population had 30 samples, and a total of 510 samples were obtained.Results: Analysis of Ae. aegypti populations in 2019 and 2020 based on microsatellite markers revealed 67 and 72 alleles, respectively. The average allelic richness of the populations in 2019 was 3.659, while that in 2020 was 3.965. The HWE analysis of the 22 populations sampled in 2019 revealed significant departure only in the QSH-2 population. The 17 populations sampled in 2020 were all in HWE. The average polymorphic information content (PIC) values were 0.546 and 0.545, respectively, showing high polymorphism. The average observed heterozygosity of the 2019 and 2020 populations was 0.538 and 0.514, respectively, and the expected average heterozygosity was 0.517 and 0.519, showing high genetic diversity in all mosquito populations. By analyzing the COI and ND4 fragments in the mitochondrial DNA of Ae. aegypti, the populations sampled in 2019 had a total of 10 COI haplotypes and 17 ND4 haplotypes. A total of 20 COI haplotypes were found in the populations sampled in 2020, and a total of 24 ND4 haplotypes were obtained. STRUCTURE, UPGMA and DAPC cluster analyses and a network diagram constructed based on COI and ND4 fragments showed that the populations of Ae. aegypti in Yunnan Province sampled in 2019 and 2020 could be divided into two clusters. At the beginning of 2020, due to the impact of COVID-19, the flow of goods between the port areas of Yunnan Province and neighboring countries was reduced, and the sterilization was more effective when goods enter the customs, leading to different immigration pressures on Ae. aegypti population in Yunnan Province between 2019 and 2020, the source populations of the 2019 and 2020 populations changed. Mantel test is generally used to detect the correlation between genetic distance and geographical distance, the analysis indicated that population geographic distance and genetic distance had a moderately significant correlation in 2019 and 2020 (2019: p < 0.05 R2 = 0.4807, 2020: p < 0.05 R2 = 0.4233).Conclusion:Ae. aegypti in Yunnan Province maintains a high degree of genetic diversity. Human interference is one reason for the changes in the genetic characteristics of this disease vector.

Background: Meniscus injuries, a common joint disease caused by long-term wear, trauma and inflammation, usually cause chronic dysfunction and pain in the joint. Current clinical surgeries mainly aim to remove the diseased tissue to alleviate patient suffering instead of helping with meniscus regeneration. As an emerging treatment, stem cell therapy has been verified to facilitate meniscus regeneration effectively. The purpose of this study is to investigate the publication conditions of stem cell therapy for meniscal regeneration and to visualize the research trends and frontiers.Methods: Relevant publications relevant to stem cells for meniscal regeneration was retrieved SCI-Expanded of the Web of Science database from 2012 to 2022. Research trends in the field were analysed and visualized by CiteSpace and VOSviewer.Results: A total of 354 publications were collected and analysed. The United States contributed the largest number of publications (118, 34.104%). Tokyo Medical Dental University has contributed the largest number of publications (34) among all full-time institutions. Stem cell research therapy has published the largest number of researches on stem cells for meniscal regeneration (17). SEKIYA. I contributed the majority of publications in this field (31), while Horie, M was the most frequently cited authors (166). #1 tissue engineering, #2 articular cartilage, #3 anterior cruciate ligament, #4 regenerative medicine, #5 scaffold are the chief keywords. This indicates that the current research hotspot has been transformed from basic surgical research to tissue engineering.Conclusion: Stem cell therapy is a promising therapeutic method for meniscus regeneration. This is the first visualized and bibliometric study to thoroughly construct the development trends and knowledge structure in the research field of stem cell therapy for meniscal regeneration in the past 10 years. The results thoroughly summarize and visualize the research frontiers, which will shed light on the research direction of stem cell therapy for meniscal regeneration.

IntroductionNumerous studies on the mosquito life cycle and transmission efficacy were performed under constant temperatures. Mosquito in wild, however, is not exposed to constant temperature but is faced with temperature variation on a daily basis.MethodsIn the present study, the mosquito life cycle and Zika virus transmission efficiency were conducted at daily fluctuating temperatures and constant temperatures. Aedes albopictus was infected with the Zika virus orally. The oviposition and survival of the infected mosquitoes and hatching rate, the growth cycle of larvae at each stage, and the infection rate (IR) of the progeny mosquitoes were performed at two constant temperatures (23°C and 31°C) and a daily temperature range (DTR, 23–31°C).ResultsIt showed that the biological parameters of mosquitoes under DTR conditions were significantly different from that under constant temperatures. Mosquitoes in DTR survived longer, laid more eggs (mean number: 36.5 vs. 24.2), and had a higher hatching rate (72.3% vs. 46.5%) but a lower pupation rate (37.9% vs. 81.1%) and emergence rate (72.7% vs. 91.7%) than that in the high-temperature group (constant 31°C). When compared to the low-temperature group (constant 23°C), larvae mosquitoes in DTR developed faster (median days: 9 vs. 23.5) and adult mosquitoes carried higher Zika viral RNA load (median log10 RNA copies/μl: 5.28 vs. 3.86). However, the temperature or temperature pattern has no effect on transovarial transmission.DiscussionThose results indicated that there are significant differences between mosquito development and reproductive cycles under fluctuating and constant temperature conditions, and fluctuating temperature is more favorable for mosquitos' survival and reproduction. The data would support mapping and predicting the distribution of Aedes mosquitoes in the future and establishing an early warning system for Zika virus epidemics.

Abstract Background Numerous systematic reviews investigating the benefit of the usage of drainage after primary total hip or knee arthroplasty have been published with divergent conclusions. We aim to determine the best available evidence and consider risk of bias of these articles and to provide recommendations. Methods A systematic search of systematic reviews published through to May 2020 was performed in MEDLINE, EMBASE and Cochrane library. Methodological quality, risk of bias and best evidence choice of included articles were evaluated by AMSTAR instrument, ROBIS tool and Jadad decision algorithm, respectively. We selected systematic reviews with high methodological quality and low risk of bias ultimately as best evidence. Results Twelve meta-analyses were included lastly. According to the ROBIS tool, seven of the included systematic reviews were with low risk of bias and five with high risk of bias. The Jadad decision algorithm suggested that two reviews conducted by Zan et al. for hip and Si et al. et al. for knee were selected as the best evidence, with highest AMSTAR score and low risk of bias. Conclusions Ten systematic reviews were included as low-quality with only two high-quality studies. Based on the current available evidence, we have insufficient confidence to draw conclusion that whether to use closed suction drainage for both total knee and hip arthroplasty. To verify the necessity and benefit of using closed suction drainage after primary total knee and hip arthroplasty, and develop exact recommendations, further studies are still required.

Fused filament fabrication (FFF), also known as fused deposition modeling (FDM™), is considered one of the most promising additive manufacturing (AM) methods for its versatility, reliability and affordability. First adopted by industries for professional uses such as rapid prototyping, then by the general public in recent years, FFF has gathered itself considerable attention. Nevertheless, despite key advancements in printer technologies and filament materials, the fabrication of robust, performing and functional parts for high-demanding practical applications remains a significant challenge. Due to intrinsic deficiencies, such as the presence of voids and weak layer-to-layer adhesion, FFF-printed parts are plagued by weak and anisotropic mechanical properties in contrast to their conventionally manufactured counterparts. With the increasing demand for designable porous structures in the fields of biomedicine, 4D printing and lightweight cellular composites, understanding the challenges presented by void presence has become more relevant than ever. As existing literature has reviewed the significance of interlayer bonding, this review focuses on documenting recent insights on the formation of voids by its categorization, research method and mechanism. The primary objective is to provide a comprehensive understanding of the two current primary methods of void research—quantitative analysis and imaging. Detailed discussions on the effects of feedstock and printing parameters on void formation are also presented. Lastly, this review discusses gaps in the current research and outlines unaddressed challenges regarding void formation and its relation with the mechanical performance of FFF parts.

Bone regeneration strategies based on mesenchymal stem cell (MSC) therapy have received widespread attention. Although MSC incorporation into bone scaffolds can help with the repair process, a large number of studies demonstrate variable effects of MSCs with some noting that the inclusion of MSCs does not provide better outcomes compared to unseeded scaffolds. This may in part be related to low cell survival following implantation and/or limited ability to continue with osteogenic differentiation for pre-differentiated cells. In this study, we incorporated MSCs into gelatin microcryogels to form microtissues, and subjected these microtissues to osteogenic induction. We then mixed as-formed microtissues with those subjected to 6 days of osteogenic induction in different ratios, and investigated their ability to induce in vitro and in vivo osteogenesis during self-assembly. Using a full-thickness rat calvarial defect model, we found that undifferentiated and osteogenically induced microtissues mixed in a ratio of 2:1 produced the best outcomes of bone regeneration. This provides a new, customizable cell-based therapeutic strategy for in vivo repair of bone defects.

BackgroundWolbachia is gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Recent reports reveal the natural infection of Wolbachia in Aedes Aegypti in Malaysia, India, Philippines, Thailand and the United States. At present, none of Wolbachia natural infection in Ae. aegypti has been reported in China.MethodsA total of 480 Ae. aegypti adult mosquitoes were collected from October and November 2018 based on the results of previous investigations and the distribution of Ae. aegypti in Yunnan. Each individual sample was processed and screened for the presence of Wolbachia by PCR with wsp primers. Phylogenetic trees for the wsp gene was constructed using the neighbour-joining method with 1,000 bootstrap replicates, and the p-distance distribution model of molecular evolution was applied.Results24 individual adult mosquito samples and 10 sample sites were positive for Wolbachia infection. The Wolbachia infection rate (IR) of each population ranged from 0 - 41.7%. The infection rate of group A alone was 0%-10%, the infection rate of group B alone was 0%-7.7%, and the infection rate of co-infection with A and B was 0-33.3%.ConclusionsWolbachia infection in wild Ae. aegypti in China is the first report based on PCR amplification of the Wolbachia wsp gene. The Wolbachia infection is 5%, and the wAlbA and wAlbB strains were found to be prevalent in the natural population of Ae. aegypti in Yunnan Province.

BackgroundMacrophages significantly contributes to symptomology and structural progression of osteoarthritis (OA) and raise increasing attention in the relative research field. Recent studies have shown that tremendous progress has been made in the research of macrophages associated with osteoarthritis. However, a comprehensive bibliometric analysis is lacking in this research field. This study aimed to introduce the research status as well as hotspots and explore the field of macrophages research in OA from a bibliometric perspective.MethodsThis study collected 1481 records of macrophages associated with osteoarthritis from 1991 to 2021 in the web of science core collection (WoSCC) database. CiteSpace, VOSviewer, and R package “bibliometrix” software were used to analyze regions, institutions, journals, authors, and keywords to predict the latest trends in macrophages associated with osteoarthritis research.ResultsThe number of publications related to macrophages associated with osteoarthritis is increasing annually. China and the USA, contributing more than 44% of publications, were the main drivers for research in this field. League of European Research Universities was the most active institution and contributed the most publications. Arthritis and Rheumatism is the most popular journal in this field with the largest publications, while Osteoarthritis and Cartilage is the most co-cited journal. Koch AE was the most prolific writer, while Bondeson J was the most commonly co-cited author. “Rheumatology”, “Orthopedics”, and “Immunology” were the most widely well-represented research areas of OA associated macrophages. “Rheumatoid arthritis research”, “clinical symptoms”, “regeneration research”, “mechanism research”, “pathological features”, and “surgery research” are the primary keywords clusters in this field.ConclusionThis is the first bibliometric study comprehensively mapped out the knowledge structure and development trends in the research field of macrophages associated with osteoarthritis in recent 30 years. The results comprehensively summarize and identify the research frontiers which will provide a reference for scholars studying macrophages associated with osteoarthritis.

Abstract Background Zika virus (ZIKV) is transmitted to humans primarily by Aedes aegypti. Previous studies on Ae. aegypti from Jiegao (JG) and Mengding (MD) in Yunnan province, China have shown that these mosquitoes are able to transmit ZIKV to their offspring through vertical transmission, indicating that these two Ae. aegypti strains pose a potential risk for ZIKV transmission. However, the vector competence of these two Ae. aegypti strains to ZIKV has not been evaluated and the molecular mechanisms influencing vector competence are still unclear. Methods Aedes aegypti mosquitoes from JG and MD were orally infected with ZIKV, and the infection rate (IR), dissemination rate (DR), transmission rate (TR) and transmission efficiency (TE) of these two mosquito strains were explored to evaluate their vector competence to ZIKV. On 2, 4 and 6 days post-infection (dpi), the small RNA profiles between ZIKV-infected and non-infected Ae. aegypti midgut and salivary gland tissues were compared to gain insights into the molecular interactions between ZIKV and Ae. aegypti. Results There were no significant differences in the IR, DR, TR and TE between the two Ae. aegypti strains (P > 0.05). However, ZIKV RNA appeared 2 days earlier in saliva of the JG strain, which indicated a higher competence of the JG strain to transmit ZIKV. Significant differences in the microRNA (miRNA) expression profiles between ZIKV-infected and non-infected Ae. aegypti were found in the 2-dpi libraries of both the midgut and salivary gland tissues from the two strains. In addition, 27 and 74 miRNAs (|log2 fold change| > 2) were selected from the miRNA expression profiles of ZIKV-infected and non-infected midgut and salivary gland tissues from the JG and MD strains, respectively. Conclusions Our results provide novel insights into the ZIKV–mosquito interactions and build a foundation for future research on how miRNAs regulate the vector competence of mosquitoes to this arbovirus. Graphical abstract

Objective: With the development of extracellular vesicles (EVs) based on stem cells research all over the world, our present study was aiming to discover the global trends in this field.Methods: All publications related to EVs based on stem cells from 1991 to 2021 were collected from the Science Citation Index-Expanded of Web of Science Subsequently, the data were evaluated using the bibliometric methodology. In terms of visualized study, the VOS viewer software was performed to investigate the bibliographic coupling, co-citation, co-authorship, and co-occurrence trends, and last for the publication’s trends involved in the field of EVs based on stem cells.Results: A total of 8,208 publications were retrieved and the relative number of global publications and research interests were increasing every year especially in recent 5 years. China rank top one in terms of total publications, prolific authors, and funds, whereas the USA made the greatest contributions with the most total citations and highest H-index to the global research. Stem cell research therapy contributed the highest publications, whereas the journal of PLOS ONE showed the best total link strength. The Shanghai Jiao Tong University, University of California System, and Harvard University were the most contributive institutions. The global studies could be divided into six clusters as follows: cancer research, musculoskeletal system research, respiratory system research, urinary system and endocrine system research, nerve system research, and cardiovascular system research. All the directions were predicted to still hotspots in near future researches in this field.Conclusion: The total number of publications about EVs based stem cells would be increasing according to the current global trends. China and the USA was the largest contributors in this field. Further efforts should be put in the directions of cancer research, musculoskeletal system research, respiratory system research, urinary system and endocrine system research, nerve system research, as well was cardiovascular system research in this field of EVs based stem cells.

Background:Aedes albopictus is an indigenous primary vector of dengue and Zika viruses in China. Wolbachia is a gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Compared with research on the dispersion of Ae. albopictus at the macrospatial level (mainly at the country or continent level), little is known about its variation and Wolbachia infection at the microspatial level, which is essential for its management. Meanwhile, no local cases of dengue fever have been recorded in the history of Nanjing, which implies that few adulticides have been applied in the city. Thus, the present study examines how the Ae. albopictus population varies and the Wolbachia infection status of each population among microspatial regions of Nanjing City.Methods: The genetic structure of 17 Aedes albopictus populations collected from urban, urban fringe, and rural regions of Nanjing City was investigated based on 9 microsatellite loci and the mitochondrial coxI gene. The Wolbachia infection status of each population was also assessed with Wolbachia A- and Wolbachia B-specific primers.Results: Nine out of 58 tested pairs of microsatellite markers were highly polymorphic, with a mean PIC value of 0.560, and these markers were therefore chosen for microsatellite genotyping analysis. The Na value of each Ae. albopictus population was very high, and the urban area populations (7.353 ± 4.975) showed a lower mean value than the urban fringe region populations (7.866 ± 5.010). A total of 19 coxI haplotypes were observed among 329 Ae. albopictus individuals via haplotype genotyping, with the highest diversity observed among the urban fringe Ae. albopictus populations (Hd = 0.456) and the lowest among the urban populations (Hd = 0.277). Each Ae. albopictus population showed significant departure from HWE, and significant population expansion was observed in only three populations from the urban (ZSL), urban fringe (HAJY), and rural areas (HSZY) (p < 0.05). Combined with DAPC analysis, all the Ae. albopictus populations were adequately allocated to two clades with significant genetic differences according to population structure analysis, and the best K value was equal to two. AMOVA results showed that most (96.18%) of the genetic variation detected in Ae. albopictus occurred within individuals (FIT = 0.22238, p < 0.0001), while no significant positive correlation was observed via isolation by distance (IBD) analysis (R2 = 0.03262, p = 0.584). The TCS network of all haplotypes showed that haplotype 1 (H1) and haplotype 4 (H4) were the most frequent haplotypes among all populations, and the haplotype frequency significantly increased from urban regions (36.84%) to rural regions (68.42%). Frequent migration was observed among Ae. albopictus populations from rural to urban regions via the urban fringe region, with four direct migration routes between rural and urban regions. Furthermore, Wolbachia genotyping results showed that most of the individuals of each population were coinfected with Wolbachia A and Wolbachia B. The independent infection rate of Wolbachia A was slightly higher than that of Wolbachia B, and no significant differences were observed among different regions.Conclusion: In the microspatial environment of Nanjing City, the urban fringe region is an important region for the dispersion of Ae. albopictus populations between rural and urban areas, and Wolbachia A and Wolbachia B coinfection is the most common Wolbachia infection status in all Ae. albopictus populations among different regions.

Mosquito-borne diseases are still threats to public health in the Zhejiang province of China. Surveillance of mosquitoes and the mosquito-borne pathogen is a vital approach for early warning, prevention, and control of the infectious disease. In this study, from 2018 to 2020, a total of 141607 female mosquitoes were caught by means of the light trap method. The main species were Culex pipiens quinquefasciatus/pallens (41.32%), Culex tritaeniorhynchus (47.6%), Aedes albopictus (2.5%), Anopheles sinensis (5.87%), Armigeres subalbatus (2.64%) and other mosquito species (0.07%). Cx. pipiens s.l. were the dominant species in two urban habitats and rural residential areas while Cx. tritaeniorhynchus was the main dominant species in the rural livestock sheds. In terms of seasonal fluctuation, Cx. pipiens s.l fluctuated at a high level from May to October. The peaks of Cx. tritaeniorhynchus, An. sinensis and Ar. subalbatus were in July. In addition, a total of 693 Ae. albopictus were collected with Biogents Mosquitaire CO2 traps in emergency surveillance of dengue fever (DF) and screened for dengue virus infection. There were three circumstances of collection: The first: the sampling time before mosquito control during the local outbreak of DF in Lucheng of Wenzhou, 2019; The second circumstance: the sampling time after mosquito control during the local outbreak of DF of other cities in 2018-2019; The third circumstance: past DF epidemic areas the sampling time before mosquito control during the local outbreak of DF in Lucheng, Wenzhou, Zhejiang, 2019. The pools formed by mosquitoes collected in these three circumstances were 3 (6.1%), 35 (71.5%), and 11 (22.4%) respectively. Of the 49 pools tested, only one in the first circumstance was positive. The full-length dengue virus sequence of ZJWZ/2019 was obtained by sequencing and uploaded to the NCBI as number OK448162. Full-length nucleotide and amino acid homology analyses showed that ZJWZ2019 and Wenzhou DF serum isolates ZJWZ-62/2019 (MW582816) and ZJWZ-18/2019 (MW582815) had the highest homology. The analysis of full genome and E gene phylogenetic trees showed that ZJWZ2019 belonged to serotype 1, genotype I, lineage II, which was evolutionarily related to OK159963/Cambodia/2019. It implies that ZJWZ2019 originated in Cambodia. This study showed the species composition, seasonal dynamics of mosquitoes in different habitats in Zhejiang province and confirmed the role of Ae. albopictus in the transmission cycle of in outbreak of DF in the Lucheng district of Wenzhou in 2019, suggesting the importance of monitoring of vector Aedes infected dengue virus in the prevention and control of DF.

Zika virus (ZIKV) has emerged as a globally important arbovirus. The virus is primarily transmitted to humans through the bite of an infective Aedes albopictus in temperate area. Vertical transmission of ZIKV by Ae. albopictus is determined and has been suggested to be a means by which the virus could persist in nature. Ae. albopictus undergoes a well-characterized photoperiodic diapause. Viruses are harbored by overwintering mosquitoes in diapause that contributes to the resurgence of vertebrate diseases in the following spring, yet little is known about the impact of diapause on the regulation of viral replication and survival. The purpose of this study is to determine that Ae. albopictus in Beijing are highly susceptible to ZIKV (92.3%), and viable virus is passed to their organs of progeny via vertical transmission. Moreover, diapause eggs (diapause incidence 97.8%) had significantly lower minimum infection rates and filial infection rates of the first gonotrophic cycle than those of the second gonotrophic cycle in the short-day photoperiod group. Regarding the development of diapause eggs, the minimum infection rates and ZIKV RNA copy number increased significantly, suggesting that virus RNA replication occurred in the diapause eggs. Meanwhile, eggs from the ZIKV-infected mosquitoes had a significantly lower hatching rate compared with uninfected mosquitoes, implying an intriguing interaction between diapause eggs and virus. The findings here suggest that vertical transmission of ZIKV from diapause eggs to progeny may have a critical epidemiological role in the dissemination and maintenance of ZIKV circulating in the vector.

Currently, modified biochar has been successfully used in the remediation of soil polluted with heavy metals. However, the effects of the modified biochar on pesticides (such as simazine) are still unclear. Herein, the environmental fate of simazine, such as decomposition, leaching, and adsorption in unamended soil, in the soil amended with unmodified and modified biochar (biochar + FeCl3, biochar + FeOS, biochar + Fe) were evaluated. In addition, an incubation experiment was also performed to observe the influence of modified biochar on the microbial community and diversity in the soil. The results showed that modified biochar significantly decreased the decomposition of simazine in the soil compared to its counterpart. Modified biochar also reduced the concentration of simazine in the leachate. Compared with the control, soil microbial biomass in the soil amended with unmodified biochar, biochar + FeCl3, biochar + Fe, and biochar + FeOS was decreased by 5.3%, 18.8%, 8.7%, and 18.1%, respectively. Furthermore, modified biochar changed the structure of the microbial community. This shows that modified biochar could increase the soil adsorption capacity for simazine and change the amount and microbial community that regulates the fate of simazine in the soil. This study concludes that iron-modified biochar has positive and negative effects on the soil. Therefore, its advantages and side effects should be considered before applying it to the soil.

Abstract The application of biochar to land has been promoted as a strategy for sequestering carbon in soils, for improving soil fertility and remediating soil pollution. However, the implications of biochar amendments on mycorrhizal associations and pesticide decomposition in agricultural soils are poorly understood. In this study, we compared the effects of four treatments; control (no biochar and no arbuscular mycorrhizal fungi (AMF), biochar (biochar without AMF), AMF (AMF without biochar) and biochar + AMF (AMF and biochar) on the fate of simazine. We specifically focused on the sorption, leaching and biodegradation behaviour of simazine. Our results showed that when symbiosis existed between plants and AMF, biochar inhibited simazine decomposition and AMF inoculation alleviated this inhibition. In contrast, this alleviation was not observed when the plant was removed. In addition, AMF inoculated into the biochar amended soil significantly decreased simazine concentration in the leachate; however, in the AMF‐only treatment, no effect on simazine leaching was observed. These phenomena were attributed to variation in the soil's sorption capacity due to biochar application or AMF inoculation. Overall, biochar application combined with AMF inoculation has the potential to mitigate simazine accumulation in the topsoil and reduce its availability.

Bioremediation is an effective way to remove heavy metals from pollutants. This study investigated the effects of Yarrowia lipolytica (Y. lipolytica) on the bioremediation of chromated copper arsenate (CCA)-treated wood wastes. Copper ions stressed the yeast strains to improve their bioremediation efficiency. A comparison of changes in morphology, chemical composition, and metal content of CCA wood before and after bioremediation was conducted. The amount of arsenic (As), chromium (Cr), and copper (Cu) was quantified by microwave plasma atomic emission spectrometer. The results showed that yeast strains remained on the surface of CCA-treated wood after bioremediation. The morphologies of the strains changed from net to spherical because of the Cu2+ stress. Fourier-transform infrared spectroscopy showed that carboxylic acid groups of wood were released after removing heavy metals. A large amount of oxalic acid was observed when the optical density (OD600nm) was 0.05 on the 21st day. Meanwhile, the highest removal rate of Cu, As, and Cr was 82.8%, 68.3%, and 43.1%, respectively. Furthermore, the Cu removal from CCA-treated wood increased by about 20% after Cu2+ stress. This study showed that it is feasible to remove heavy metals from CCA-treated wood by Y. lipolytica without destroying the wood structure, especially by copper-induced Y. lipolytica.

Difenoconazole is a type of triazole fungicide that is widely used in the treatment of plant diseases. Triazole fungicides have been shown in several studies to impair the development of the nervous system in zebrafish embryos. There is still little known about difenoconazole-induced neurotoxicity in fish. In this study, zebrafish embryos were exposed to 0.25, 0.5, and 1 mg/L of difenoconazole solution until 120 h post-fertilization (hpf). The difenoconazole-exposed groups showed concentration-dependent inhibitory tendencies in heart rate and body length. Malformation rate and spontaneous movement of zebrafish embryos increased, and the locomotor activity decreased in the highest exposure group. The content of dopamine and acetylcholine was reduced significantly in difenoconazole treatment groups. The activity of acetylcholinesterase (AChE) was also increased after treatment with difenoconazole. Furthermore, the expression of genes involved in neurodevelopment was remarkably altered, which corresponded with the alterations of neurotransmitter content and AChE activity. These results indicated that difenoconazole might affect the development of the nervous system through influencing neurotransmitter levels, enzyme activity, and the expression of neural-related genes, ultimately leading to abnormal locomotor activity in the early stages of zebrafish.

Abstract Background Aedes albopictus is an indigenous primary vector for dengue and Zika viruses in China. Compared with its insecticide resistance, biology and vector competence, little is known about its genetic variation, which corresponds to environmental variations. Thus, the present study examines how Ae. albopictus varies among different climatic regions in China and deciphers its potential dispersal patterns. Methods The genetic variation and population structure of 17 Ae. albopictus populations collected from three climatic regions of China were investigated with 11 microsatellite loci and the mitochondrial coxI gene. Results Of 44 isolated microsatellite markers, 11 pairs were chosen for genotyping analysis and had an average PIC value of 0.713, representing high polymorphism. The number of alleles was high in each population, with the n e value increasing from the temperate region (3.876) to the tropical region (4.144). Twenty-five coxI haplotypes were detected, and the highest diversity was observed in the tropical region. The mean Ho value (ca. 0.557) of all the regions was significantly lower than the mean He value (ca. 0.684), with nearly all populations significantly departing from HWE and displaying significant population expansion (p value 0.4) were reconstructed between the tropical region and the other two regions. Environmental factors, especially temperature and rainfall, may be the leading causes of genetic diversity in different climatic regions. Conclusions Continuous dispersion contributes to the genetic communication of Ae. albopictus populations across different climatic regions, and environmental factors, especially temperature and rainfall, may be the leading causes of genetic variation. Graphical abstract