Genomic divergence and mutation load in the Begonia masoniana complex from limestone karsts.

Understanding genome-wide diversity, inbreeding, and the burden of accumulated deleterious mutations in small and isolated populations is essential for predicting and enhancing population persistence and resilience. However, these effects are rarely studied in limestone karst plants. Here, we re-sequenced the nuclear genomes of 62 individuals of the Begonia masoniana complex (B. liuyanii, B. longgangensis, B. masoniana and B. variegata) and investigated genomic divergence and genetic load for these four species. Our analyses revealed four distinct clusters corresponding to each species within the complex. Notably, there was only limited admixture between B. liuyanii and B. longgangensis occurring in overlapping geographic regions. All species experienced historical bottlenecks during the Pleistocene, which were likely caused by glacial climate fluctuations. We detected an asymmetric historical gene flow between group pairs within this timeframe, highlighting a distinctive pattern of interspecific divergence attributable to karst geographic isolation. We found that isolated populations of B. masoniana have limited gene flow, the smallest recent population size, the highest inbreeding coefficients, and the greatest accumulation of recessive deleterious mutations. These findings underscore the urgency to prioritize conservation efforts for these isolated population. This study is among the first to disentangle the genetic differentiation and specific demographic history of karst Begonia plants at the whole-genome level, shedding light on the potential risks associated with the accumulation of deleterious mutations over generations of inbreeding. Moreover, our findings may facilitate conservation planning by providing critical baseline genetic data and a better understanding of the historical events that have shaped current population structure of rare and endangered karst plants.

Phylogeographic patterns match the floristic subdivisions: The diversification history of a widespread herb in subtropical China.

BACKGROUND AND AIMS: Subtropical China is dominated by evergreen broad-leaved forests (EBLFs) and is acknowledged as a critical region for its high floristic richness and endemism. Understanding of evolutionary mechanisms of such global biodiversity hotspots comes almost exclusively from long-lived tree species. Herbaceous plants represent critical biodiversity components in forests, however, the diversification history of understory herbs in subtropical EBLFs remain poorly understood. Here, we investigated the phylogeographic patterns and demographic history of Oreocharis auricula, a widespread perennial herb endemic to the EBLFs of subtropical China. METHODS: Both cpDNA sequences and single-copy nuclear genes were used to investigate the genetic variation among 657 individuals from 68 populations. Evidences from molecular dating, demographic history construction, and species distribution modeling were also combined to infer the phylogeography and evolutionary history of O. auricula. KEY RESULTS: Strong phylogeographic signals have been congruently observed using nuclear and plastid DNA markers, with the diversification patterns generally consistent with the recognized floristic subdivisions of subtropical China. Notably, we revealed an important phylogeographic barrier along the Nanling mountain range, which is also around a climatic transition at 24-26°N latitude in subtropical China, separating the south monsoon subtropical EBLFs from the mid-subtropical EBLFs. Demographic expansion and significant niche divergence were detected among the extant lineages, which may have diverged during the early Pleistocene. CONCLUSIONS: The inherent characteristics of understory herbs with limited dispersal and short generation time intensify the genetic divergence response of O. auricula to abiotic forces, contributing to the profound phylogeographic imprints of mountains and climate in such herbaceous flora. To further substantiate the generality of the identified patterns, it is paramount to extend phylogeographic investigations to other understory herbaceous taxa in subtropical China. These results have expanded our understanding of the diversification processes of subtropical forests in China.

The impact of bloodmeal and geographic region on the richness, diversity, and function of internal microbial community in Haemaphysalis qinghaiensis from the Qinghai province, China.

Background: Ticks are ectoparasites that feed on blood and pose a threat to both the livestock industry and public health due to their ability to transmit pathogens through biting. However, the impact of factors such as bloodmeal and geographic regions on the bacterial microbiota of Haemaphysalis qinghaiensis remains poorly understood. Methods: In this study, we used the v3-v4 region of the 16S rRNA gene to sequence the microbiota of Haemaphysalis qinghaiensis from eight groups (HY_M, YS_M, XH_M, LD_M, BM_M, LD_F_F, LD_F, and BM_F_F) in Qinghai Province. Results: Significant differences in bacterial richness were observed between LD_F_F, BM_F_F, and LD_F (P < 0.01), and among the five groups (HY_M, YS_M, XH_M, BM_M, and LD_M) (P < 0.05). The bacterial diversity also differed significantly between LD_F_F, LD_F, and BM_F_F (P < 0.01), as well as among the five groups (HY_M, YS_M, XH_M, LD_M, and BM_M) (P < 0.01). The group with the highest number of operational taxonomic units (OTUs) was LD_F, accounting for 23.93 % (419/1751), while BM_F_F accounted for at least 0.80 % (14/1751). At the phylum level, Firmicutes was the most abundant, with relative abundance ranging from 7.44 % to 96.62 %. At the genus level, Staphylococcus had the highest abundance, ranging from 1.67 % to 97.53 %. The endosymbiotic bacteria Coxiella and Rickettsia were predominantly enriched in LD_F_F. Additionally, the 16S gene of Coxiella showed the highest identity of 99.07 % with Coxiella sp. isolated from Xinxiang hl9 (MG9066 71.1), while the 16S gene of Rickettsia had 100 % identity with Candidatus Rickettsia hongyuanensis strains (OK 662395.1). Functional predictions for the prokaryotic microbial community indicated that the main functional categories were Metabolic, Genetic information processing, and Environmental information processing across the eight groups. Conclusion: This study provides a theoretical basis for the prevention and treatment of tick-borne diseases, which is of great significance for public health.

Correlation between change in serum creatinine concentration and renal cortical anisotropic backscattering artifact in azotemic cats during hospitalization.

Information on the clinical outcomes of feline azotemia using ultrasound examinations is limited. This study aimed to understand the correlation between cortical anisotropy backscattering artifact (CABA) and serum creatinine (sCr) changes in feline azotemia after hospitalization and to investigate whether CABA is useful for predicting the clinical outcome of feline azotemia. Sixty-five hospitalized cats with azotemia, including 49 cats with moderate or severe azotemia (severe group) and 16 cats with mild azotemia (mild group). This retrospective study reviewed the CABA using ultrasound images of cats hospitalized with azotemia between 2016 and 2021. The correlation between CABA and the clinical outcomes of cats with azotemia was investigated using the chi-squared or Fisher's exact test, and the intra- and inter-observer agreements in CABA were assessed using McNemar's and Cohen's kappa tests. The presence of CABA was significantly positively correlated with the clinical outcomes of cats with azotemia only in the severe group (p = 0.0034, odds ratio = 8.57). There was no association between CABA and clinical outcomes in cats with mild azotemia (p = 0.75). CABA can be used for clinical outcome prediction in moderate and severe feline azotemia, with a sensitivity of 80.8% and a specificity of 73.9%. Also, satisfactory intra- and inter-observer agreements were revealed in the detection of CABA during ultrasound image review. Our study demonstrated that cats with moderate and severe azotemia with CABA observed during ultrasonography might have better clinical outcomes. These findings provide additional information on the prognosis and treatment of feline azotemia.

Application of predictive model based on CT radiomics and machine learning in diagnosis for occult locally advanced esophageal squamous cell carcinoma before treatment: A two-center study.

PURPOSE: Development and validation of a radiomics model for predicting occult locally advanced esophageal squamous cell carcinoma (LA-ESCC) on computed tomography (CT) radiomic features before implementation of treatment. METHODS: The study retrospectively collected 574 patients with esophageal squamous cell carcinoma (ESCC) from two medical centers, which were divided into three cohorts for training, internal and external validation. After delineating volume of interest (VOI), radiomics features were extracted and subjected to feature selection using three robust methods. Subsequently, 10 machine learning models were constructed, among which the optimal model was utilized to establish a radiomics signature. Furthermore, a predictive nomogram incorporating both clinical and radiomics signatures was developed. The performance of these models was evaluated through receiver operating characteristic curves, calibration curves, decision curve analysis as well as measures including accuracy, sensitivity, and specificity. RESULTS: A total of 19 radiomics features were selected. The multilayer perceptron (MLP), which was found to be optimal, achieved an AUC of 0.919, 0.864 and 0.882 in the training, internal and external validation cohorts, respectively. Similarly, MLP showed good accuracy in distinguish occult LA-ESCC in subgroup of cT1-2N0M0 diagnosed by clinicians with 0.803 and 0.789 in two validation cohorts respectively. By incorporating the radiomics signature with clinical signature, a predictive nomogram demonstrated superior prediction performance with an AUC of 0.877 and accuracy of 0.85 in external validation cohort. CONCLUSION: The radiomics and machine learning model can offers improved accuracy in prediction of occult LA-ESCC, providing valuable assistance to clinicians when choosing treatment plans.

Pseudo coherent-perfect-absorption approach toward perfect polarization conversion.

Polarization is one of the essential properties of light. Thereby, its manipulation is important for numerous applications. When employing a resonance in a mirror-symmetry system to manipulate polarization, non-zero residual light in the excited polarization channel leads to the shrink in the scope of the polarization manipulation, and a perfect polarization conversion cannot occur. In this work we show that the concept of coherent perfect absorption can be applied to perfect polarization conversion for circular polarization states. We find that the only requirement to achieve a perfect polarization conversion is that the working frequency is the resonant one. More importantly, the range of the output polarization states can be efficiently enlarged, and can span the entire Poincare sphere by combining the momentum dependent radiative coupling rate driven by the bound states in the continuum (BIC) and the phase delay. When applied to realistic design, we adopt a guided mode resonance driven from the symmetry protected BICs in a dielectric photonic crystal slab. Numerical results are in good agreements with our theoretical predictions. We believe this work can deliver important benefits for a variety of applications based on the efficiently light polarization control and management.

Clinically relevant immune subtypes based on alternative splicing landscape of immune-related genes for lung cancer advanced PPPM approach.

Background: Alternative splicing (AS) occurs in the process of gene post-transcriptional process, which is very important for the correct synthesis and function of protein. The change of AS pattern may lead to the change of expression level or function of lung cancer-related genes, and then affect the occurrence and development of lung cancers. The specific AS pattern might be used as a biomarker for early warning and prognostic assessment of a cancer in the framework of predictive, preventive, and personalized medicine (PPPM; 3PM). AS events of immune-related genes (IRGs) were closely associated with tumor progression and immunotherapy. We hypothesize that IRG-AS events are significantly different in lung adenocarcinomas (LUADs) vs. controls or in lung squamous cell carcinomas (LUSCs) vs. controls. IRG-AS alteration profiling was identified to construct IRG-differentially expressed AS (IRG-DEAS) signature models. Study on the selective AS events of specific IRGs in lung cancer patients might be of great significance for further exploring the pathogenesis of lung cancer, realizing early detection and effective monitoring of lung cancer, finding new therapeutic targets, overcoming drug resistance, and developing more effective therapeutic strategies, and better used for the prediction, diagnosis, prevention, and personalized medicine of lung cancer. Methods: The transcriptomic, clinical, and AS data of LUADs and LUSCs were downloaded from TCGA and its SpliceSeq databases. IRG-DEAS events were identified in LUAD and LUSC, followed by their functional characteristics, and overall survival (OS) analyses. OS-related IRG-DEAS prognostic models were constructed for LUAD and LUSC with Lasso regression, which were used to classify LUADs and LUSCs into low- and high-risk score groups. Furthermore, the immune cell distribution, immune-related scores, drug sensitivity, mutation status, and GSEA/GSVA status were analyzed between low- and high-risk score groups. Also, low- and high-immunity clusters and AS factor (SF)-OS-related-AS co-expression network and verification of cell function of CELF6 were analyzed in LUAD and LUSC. Results: Comprehensive analysis of transcriptomic, clinical, and AS data of LUADs and LUSCs identified IRG-AS events in LUAD (n = 1607) and LUSC (n = 1656), including OS-related IRG-AS events in LUAD (n = 127) and LUSC (n = 105). A total of 66 IRG-DEAS events in LUAD and 89 IRG-DEAS events in LUSC were identified compared to controls. The overlapping analysis between IRG-DEASs and OS-related IRG-AS events revealed 14 OS-related IRG-DEAS events for LUAD and 16 OS-related IRG-DEAS events for LUSC, which were used to identify and optimize a 12-OS-related-IRG-DEAS signature prognostic model for LUAD and an 11-OS-related-IRG-DEAS signature prognostic model for LUSC. These two prognostic models effectively divided LUAD or LUSC samples into low- and high-risk score groups that were closely associated with OS, clinical characteristics, and tumor immune microenvironment, with significant gene sets and pathways enriched in the two groups. Moreover, weighted gene co-expression network (WGCNA) and nonnegative matrix factorization method (NMF) analyses identified four OS-relevant subtypes of LUAD and six OS-relevant subtypes of LUSC, and ssGSEA identified five immunity-relevant subtypes of LUAD and five immunity-relevant subtypes of LUSC. Interestingly, splicing factors-OS-related-AS network revealed hub molecule CELF6 was significantly related to the malignant phenotype in lung cancer cells. Conclusions: This study established two reliable IRG-DEAS signature prognostic models and constructed interesting splicing factor-splicing event networks in LUAD and LUSC, which can be used to construct clinically relevant immune subtypes, patient stratification, prognostic prediction, and personalized medical services in the PPPM practice. Supplementary Information: The online version contains supplementary material available at 10.1007/s13167-024-00366-4.

Chromosome-level genome assembly of Euphorbia tirucalli (Euphorbiaceae), a highly stress-tolerant oil plant.

Euphorbia, one of the largest genera of flowering plants, is well-known for containing many biofuel crops. Euphorbia tirucalli, an evergreen succulent mainly native to the Africa continent but cultivated worldwide, is a promising petroleum plant with high tolerance to drought and salt stress. However, the exploration of such an important plant resource is severely hampered by the lack of a reference genome. Here, we present the chromosome-level genome assembly of E. tirucalli using PacBio HiFi sequencing and Hi-C technology. Its genome size was approximately 745.62 Mb, with a contig N50 of 74.16 Mb. A total of 743.63 Mb (99.73%) of the assembled sequences were anchored to 10 chromosomes with a complete BUSCO score of 97.80%. Genome annotation revealed 26,304 protein-coding genes, and 76.37% of the genome was identified as repeat elements. The high-quality genome provides valuable genetic resources that would be useful for unraveling the genetic mechanisms of biofuel synthesis and evolutionary adaptation of E. tirucalli.

Demonstration of π-π Stacking at Interfaces: Synthesis of an Indole-Modified Monodisperse Silica Microsphere SiO2@IN.

In the present study, a novel silane coupling agent, designated INSi, was synthesized via a facile synthetic route, incorporating indole-functional moieties. This agent was further employed for the surface modification of homemade silica nanomicrospheres (SMPs). The ensuing nanomicrosphere composite, denoted as SiO2@IN, exemplified pronounced interfacial π-π interactions. Optimization of the reaction conditions was conducted using the response surface optimization technique. Subsequent validation of interfacial π-π interactions was accomplished through a synergistic approach, integrating theoretical calculations and comprehensive analyses of spectral and morphological attributes exhibited by the SiO2@IN.

Whether Patients with Anterior Cruciate Ligament Reconstruction Walking at a Fast Speed Show more Kinematic Asymmetries?

OBJECTIVE: Knee kinematic asymmetries after anterior cruciate ligament reconstruction (ACLR) are correlated with poor clinical outcomes, such as the progression of knee cartilage degenerations or reinjuries. Fast walking in patients with knee conditions may exacerbate knee kinematic asymmetries, but its impact on ACLR patients is uncertain. The aim of this study is to investigate if fast walking induces more knee kinematic asymmetries in unilateral ACLR patients. METHODS: This cross-sectional study enrolled 55 patients with unilateral ACLR from January 2020 to July 2022. There were 48 males and seven females with an average age of 30.6 ± 6.4 years. Knee kinematic data were collected at three walking speeds: self-selected, fast (150% normal), and slow (50% normal). A 3D knee kinematic analysis system measured the data, and self-reported outcomes assessed comfort levels during walking. We used SPM1D for two-way repeated ANOVA and posthoc paired t-tests to analyze kinematic differences in groups. RESULTS: In fast walking, ACLR knees exhibited more transverse kinematic asymmetries than intact knees, including greater external rotation angle (1.8°, 38%-43%; gait cycle [GC], p < 0.05 & 1.8-2.7°, 50%-61% GC, p < 0.05) and increased proximal tibial translation (2.1-2.5 mm, 2%-6% GC, p < 0.05 & 2.5-3.2 mm, 92%-96% GC, p < 0.05). Additionally, ACLR knees showed greater posterior tibial translation than intact knees (3.6-3.7 mm, 7%-8% GC, p < 0.05) during fast walking. No posterior tibial translation asymmetries were observed in slow walking compared to normal walking levels. ACLR knees have the most comfortable feelings in slow walking speed, and the most uncomfortable feelings in fast walking speed levels (29%). CONCLUSIONS: Fast walking induces additional external tibial rotation and proximal and posterior tibial translation asymmetries in ACLR patients. This raises concerns about long-term safety and health during fast walking. Fast walking, not self-selected speed, is beneficial for identifying postoperative gait asymmetries in ACLR patients.

Adjuvant therapy provides no additional recurrence-free benefit for esophageal squamous cell carcinoma patients after neoadjuvant chemoimmunotherapy and surgery: a multi-center propensity score match study.

Purpose: The need for adjuvant therapy (AT) following neoadjuvant chemoimmunotherapy (nICT) and surgery in esophageal squamous cell cancer (ESCC) remains uncertain. This study aims to investigate whether AT offers additional benefits in terms of recurrence-free survival (RFS) for ESCC patients after nICT and surgery. Methods: Retrospective analysis was conducted between January 2019 and December 2022 from three centers. Eligible patients were divided into two groups: the AT group and the non-AT group. Survival analyses comparing different modalities of AT (including adjuvant chemotherapy and adjuvant chemoimmunotherapy) with non-AT were performed. The primary endpoint was RFS. Propensity score matching(PSM) was used to mitigate inter-group patient heterogeneity. Kaplan-Meier survival curves and Cox regression analysis were employed for recurrence-free survival analysis. Results: A total of 155 nICT patients were included, with 26 patients experiencing recurrence. According to Cox analysis, receipt of adjuvant therapy emerged as an independent risk factor(HR:2.621, 95%CI:[1.089,6.310], P=0.032), and there was statistically significant difference in the Kaplan-Meier survival curves between non-AT and receipt of AT in matched pairs (p=0.026). Stratified analysis revealed AT bring no survival benefit to patients with pathological complete response(p= 0.149) and residual tumor cell(p=0.062). Subgroup analysis showed no significant difference in recurrence-free survival between non-AT and adjuvant chemoimmunotherapy patients(P=0.108). However, patients receiving adjuvant chemotherapy exhibited poorer recurrence survival compared to non-AT patients (p= 0.016). Conclusion: In terms of recurrence-free survival for ESCC patients after nICT and surgery, the necessity of adjuvant therapy especially the adjuvant chemotherapy, can be mitigated.

Aldehyde-free and bio-based durable coatings for cellulose fabrics with high flame retardancy, antibacteria and well wearing performance.

The bio-based coatings of cellulose fabrics (cotton) had attracted increasing attention for multifunction and sustainability but suffered from poor durability and low efficiency. Here, the aldehyde-free and durable coatings for cotton fabrics (CPZ@CF) with satisfactory flame retardancy, antibacteria as well as wearing performance were prepared through the interfacial coordination effect where the well-organized zinc phytate complex were in situ grew on the pre-treated surface of cotton fabrics with chitosan (CS) and Zn2+. The CZP@CF exhibited excellent antibacterial activity for Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) with 99.99 % antibacterial rates benefiting from the synergistic effect between Zn2+ and CS. Meanwhile, even the CPZ coatings loading was only 1.5 wt%, the fire safety of CZP@CF remarkably enhanced owing to the excellent synergistic catalytic charring and free radical capture. More importantly, the antibacterial rates of CZP@CF for S. aureus and E. coli still reached 99.99 % and 91.67 % after 50 washing cycles. Additionally, this treatment method did not deteriorate the fabrics properties, including mechanical and breathability as well as wearing performance, which provided the approach to fabricate the flame retardant and antibacterial textiles with well durability and wearing performance.

A chromosome-scale assembly of the early-flowering Prunus campanulata and comparative genomics of cherries.

Prunus campanulata is an important flowering cherry germplasm of high ornamental value. Given its early-flowering phenotypes, P. campanulata could be used for molecular breeding of ornamental species and fruit crops belonging to the subgenus Cerasus. Here, we report a chromosome-scale assembly of P. campanulata with a genome size of 282.6 Mb and a contig N50 length of 12.04 Mb. The genome contained 24,861 protein-coding genes, of which 24,749 genes (99.5%) were functionally annotated, and 148.20 Mb (52.4%) of the assembled sequences are repetitive sequences. A combination of genomic and population genomic analyses revealed a number of genes under positive selection or accelerated molecular evolution in P. campanulata. Our study provides a reliable genome resource, and lays a solid foundation for genetic improvement of flowering cherry germplasm.

Intradural extramedullary spinal choristoma: A case report.

RATIONALE: A choristoma is a rare and benign neoplasm characterized by the presence of normal tissue in an anomalous anatomical location. In contrast, choristoma tend to occur in other body regions rather than within the spinal canal. Before our findings, only 4 cases of intraspinal choristoma had been recorded. Because its composition is complex and very rare, routine examinations, such as magnetic resonance imaging, are difficult to diagnose, and the possibility of its occurrence is often missed in clinical diagnosis. If there is no specificity in its components, such as in this case, even pathological examinations can only confirm the diagnosis as choristoma after eliminating other possibilities. Therefore, in clinical practice, when encountering patients with intraspinal tumors, it is essential to consider the possibility of choristoma. In this case, the choristoma lack of specific constituent composition sets it apart from previously reported intraspinal choristoma, significantly raising the diagnostic challenge, which offers valuable insights for clinical diagnosis. PATIENT CONCERNS: A female patient aged 48 years was admitted to our medical center due to experiencing persistent lower back pain accompanied by radiating pain in both legs for 5 months. Based on the findings from the neurological physical examination and magnetic resonance imaging, the patient was diagnosed with an intradural space-occupying lesion located at the level of the first lumbar vertebral body. We performed an enhanced magnetic resonance neurography examination to further determine the positional relationship between the occupation and nerves in preparation for surgery. Postoperative pathological biopsy showed that the mass was an intraspinal choristoma. DIAGNOSIS: Intradural extramedullary spinal choristoma. INTERVENTION: Occupied lesion is removed surgically. OUTCOMES: After surgery, all symptoms were significantly relieved, and when the patient was discharged, all symptoms disappeared completely. There was no sign of recurrence after 1 year of follow-up. LESSONS: Intraspinal choristomas are not specific and need to be diagnosed by pathologic examination. Early detection of and intervention for intraspinal tumors can mitigate nerve dysfunction.

ATGL promotes colorectal cancer growth by regulating autophagy process and SIRT1 expression.

CRC is a common malignant tumor in the gastrointestinal tract, and its incidence has increased significantly in recent years. Several studies revealed that lipid metabolism reprogramming contributed to tumorigenicity and malignancy by interfering with energy production, membrane formation, and signal transduction in cancers. ATGL is a kind of hydroxy fatty acid ester of fatty acid synthase, and its role in tumor remains controversial. We compared levels of adipose triglyceride lipase (ATGL) in human CRC specimens to adjacent specimens. To validate the effect of ATGL on the proliferation ability of CRC, CCK8 assay and clone formation assay were performed. To evaluate whether autophagy process takes part in the effect of ATGL on CRC proliferation, the value of LC3-II/LC3-I was detected by western blot and we blocked the SIRT1 to detect value of LC3-II/LC3-I and p62 via western blot. In the end, we detected the value of SIRT1 in CRC specimens. We found that ATGL showed high expression in CRC and positively correlated with clinical stage, indicating poor prognosis of CRC. Moreover, ATGL significantly promoted tumor cell proliferation in vitro. Mechanistically, ATGL promoted CRC cells proliferation by blocking mTOR signaling pathway and activating autophagy process. Further, ATGL regulated autophagy process through triggering SIRT1 expression. Our results reveal that ATGL promotes colorectal cancer growth by up regulating autophagy process and SIRT1 expression.

First description study on molecular characterization of Hemiclepsis khankiana (Hirudinea: Glossiphoniidae) in China.

BACKGROUND: Leeches are an integral component of aquatic biocenosis and can be found in a wide range of ecosystems such as freshwater, saltwater, flowing, and still-water ecosystems. It especially plays an important role in the freshwater benthic community and is an important part of the food web. In this study, a leech species was found in the mantle cavity of wild freshwater mussels in Zigong City, Sichuan Province, China, and its identity was determined through morphological analysis and molecular biological analysis. RESULTS: The leech is Hemiclepsis khankiana, a new species of Hemiclepsis that has been discovered in Russia in recent years. Through morphological analysis, the current survey observed that the morphological characteristics of Hemiclepsis khankiana eyespots were significantly different from the first reported description. The first pair of eyespots on the leech were separated and clear, while it had been reduced to unclear shadows in the previous report. The phylogenetic tree based on the COI gene showed that the COI gene sequence obtained in this study was in the same evolutionary branch as Hemiclepsis khankiana (MN295420, MN295421). Genetically, it was most closely related to Hemiclepsis kasmiana (mean COI p-distance = 3.98%). CONCLUSIONS: The current study reported on the new distribution range of Hemiclepsis khankiana, which was initially discovered in China. This study indicates that the distribution range of the leech species has expanded, laying a foundation for further studies in China.

Hemalin vaccination modulates the host immune response and reproductive cycle of Haemaphysalis longicornis.

Haemaphysalis longicornis can transmit high varieties of tick-borne pathogens (TBPs), and a primary strategy for preventing the transmission of those TBPs is to control ticks. Hemalin, a thrombin inhibitor of the Kunitz-type family and a crucial component in H. longicornis feeding process has been isolated from parthenogentic ticks. This study aimed to evaluate the validity of a recombinant Hemalin (rHlHemalin) vaccination as an anti-tick vaccine against H. longicornis in rabbits to find a new candidate for an effective tick control. In this study, mouse splenocytes were isolated and used to investigate immune responses after rHlHemalin stimulation. The rabbits were vaccinated with the rHlHemalin protein. After tick challenges, body weight at engorgement, egg mass, and the reproductive cycle of H. longicornis were evaluated. To confirm the vaccination, the passive immunization tests of α-rHlHemalin sera were performed. The results showed that the rHlHemalin protein could stimulate cytokine production in mouse splenocytes. Vaccination assay revealed that the periods from tick infestations to egg-hatch in the vaccination group were significantly longer than those in the phosphate buffer saline (PBS) group (P = 0.0003). In addition, the tick body weight at engorgement (P = 0.0019) and egg mass at 10 days after oviposition (P = 0.0232) were higher than those in the PBS group. These findings were consistent with the current passive immunization results and suggest rHlHemalin vaccination extended the reproductive cycle in H. longicornis but did not decrease the body weight at engorgement or weight of egg mass. Therefore, it is debatable whether Hemalin vaccination is highly-effective anti-tick vaccine or not. However, due to the importance of thrombin inhibitors in tick blood feeding and blood digestion, additional inhibitor-based vaccines should be developed aiming to find an effective and environmentally friendly biological strategy to combat ticks.

The genetic structure and demographic history revealed by whole-genome resequencing provide insights into conservation of critically endangered Artocarpus nanchuanensis.

Introduction: Whole-genome resequencing technology covers almost all nucleotide variations in the genome, which makes it possible to carry out conservation genomics research on endangered species at the whole-genome level. Methods: In this study, based on the whole-genome resequencing data of 101 critically endangered Artocarpus nanchuanensis individuals, we evaluated the genetic diversity and population structure, inferred the demographic history and genetic load, predicted the potential distributions in the past, present and future, and classified conservation units to propose targeted suggestions for the conservation of this critically endangered species. Results: Whole-genome resequencing for A. nanchuanensis generated approximately 2 Tb of data. Based on abundant mutation sites (25,312,571 single nucleotide polymorphisms sites), we revealed that the average genetic diversity (nucleotide diversity, π) of different populations of A. nanchuanensis was relatively low compared with other trees that have been studied. And we also revealed that the NHZ and QJT populations harboured unique genetic backgrounds and were significantly separated from the other five populations. In addition, positive genetic selective signals, significantly enriched in biological processes related to terpene synthesis, were identified in the NHZ population. The analysis of demographic history of A. nanchuanensis revealed the existence of three genetic bottleneck events. Moreover, abundant genetic loads (48.56% protein-coding genes) were identified in Artocarpus nanchuanensis, especially in genes related to early development and immune function of plants. The predication analysis of suitable habitat areas indicated that the past suitable habitat areas shifted from the north to the south due to global temperature decline. However, in the future, the actual distribution area of A. nanchuanensis will still maintain high suitability. Discussion: Based on total analyses, we divided the populations of A. nanchuanensis into four conservation units and proposed a number of practical management suggestions for each conservation unit. Overall, our study provides meaningful guidance for the protection of A. nanchuanensis and important insight into conservation genomics research.