Inkjet printer prediction under complicated printing conditions based on microscopic image features.

A novel technique is introduced to predict the printer model used to produce a given document. Samples containing only a few letters printed under varying conditions (i.e., different printing modes, letter types, fonts) were collected to establish a dataset of 41 inkjet printer models from common manufacturers, such as HP, Canon, and Epson. Morphological features were analyzed by extraction of image features using several algorithms in a series of microscopic images and a Wilcoxon test was used to measure the significance of variations between printed samples. Significant differences between various printing conditions might post potential challenge to questioned document examination. Discriminant analysis and the k-nearest neighbor (KNN) algorithm were also employed for source printer prediction under varying printing condition on 30% images with the rest images as training dataset. The results of a validation experiment demonstrated that while quadratic discriminant analysis (QDA) achieved an accuracy of 96.3%, a combination of KNN and QDA reached 98.6%. As such, this technique could aid in the forensic examination of printed documents.

Occupational fibrotic hypersensitivity pneumonia in a halogen dishes manufacturer: A case report.

BACKGROUND: Fibrotic hypersensitivity pneumonitis (FHP) is an allergic and diffuse pneumonia caused by repeated inhalation of antigenic substances, and sometimes developed in people working in specific environments. While novel antigens and exposures continued to be described, physicians should maintain a high suspicion of potential exposures. A detailed assessment of the patient's occupational exposures as well as living environment is necessary and complete allergen avoidance is the first and most important step in the management of FHP once the allergens are determined. CASE SUMMARY: A 35-year-old female was admitted to the hospital with a cough and breathing difficulties for more than one year. She was a nonsmoker and a manufacturer of halogen dishes, which are characteristic Chinese foods, for 15 years without any protection. High resolution computed tomography of the chest demonstrated an interstitial pneumonia pattern. Pulmonary function examination showed restricted ventilation dysfunction and a significant reduction in dispersion ability. Cell differentiation in bronchoalveolar lavage fluid demonstrated lymphocytosis (70.4%) with an increased lymphocyte CD4/CD8 ratio (0.94). Transbronchial lung biopsy combined with lung puncture pathology showed diffuse uniform alveolar interval thickening, chronic inflammatory cell infiltration, a proliferation of tissue in the bronchial wall fiber and alveolar epithelial follicle degeneration, resulting in fibrosis. CONCLUSION: Exposure to spices used for the production of halogen dishes may cause FHP.

The efficacy of Raman spectroscopy in lung cancer diagnosis: the first diagnostic meta-analysis.

In recent years, many researches have explored the diagnostic value of Raman spectroscopy in multiple types of tumors. However, as an emerging clinical examination method, the diagnostic performance of Raman spectroscopy in lung cancer remains unclear. Relevant diagnostic studies published before 1 June 2020 were retrieved from the Cochrane Library, PubMed, EMBASE, China National Knowledge Internet (CNKI), and WanFang databases. After the literature was screened, two authors extracted the data from eligible studies according to the inclusion and exclusion criteria. Obtained data were pooled and analyzed using Stata 16.0, Meta-DiSc 1.4, and RevMan 5.3 software. Fourteen diagnostic studies were eligible for the pooled analysis which includes 779 patients. Total pooled sensitivity and specificity of Raman spectroscopy in diagnosing lung cancer were 0.92 (95% CI 0.87-0.95) and 0.94 (95% CI 0.88-0.97), respectively. The positive likelihood ratio was 15.2 (95% CI 7.5-30.9), the negative likelihood ratio was 0.09 (95% CI 0.05-0.14), and the area under the curve was 0.97 (95 % CI 0.95-0.98). Subgroup analysis suggested that the sensitivity and specificity of RS when analyzing human tissue, serum, and saliva samples were 0.95 (95% CI 0.88-0.98), 0.97 (95% CI 0.89-0.99), 0.88 (95% CI 0.80-0.93), 0.87 (95% CI 0.78-0.92), 0.91 (95% CI 0.80-0.96), and 0.95 (95% CI 0.73-0.99), respectively. No publication bias or threshold effects were detected in this meta-analysis. This initial meta-analysis indicated that Raman spectroscopy is a highly specific and sensitive diagnostic technology for detecting lung cancer. Further investigations are also needed to focus on real-time detection using Raman spectroscopy under bronchoscopy in vivo. Moreover, large-scale diagnostic studies should be conducted to confirm this conclusion.

Giant tortoise genomes provide insights into longevity and age-related disease.

Giant tortoises are among the longest-lived vertebrate animals and, as such, provide an excellent model to study traits like longevity and age-related diseases. However, genomic and molecular evolutionary information on giant tortoises is scarce. Here, we describe a global analysis of the genomes of Lonesome George-the iconic last member of Chelonoidis abingdonii-and the Aldabra giant tortoise (Aldabrachelys gigantea). Comparison of these genomes with those of related species, using both unsupervised and supervised analyses, led us to detect lineage-specific variants affecting DNA repair genes, inflammatory mediators and genes related to cancer development. Our study also hints at specific evolutionary strategies linked to increased lifespan, and expands our understanding of the genomic determinants of ageing. These new genome sequences also provide important resources to help the efforts for restoration of giant tortoise populations.

Hypoxia promotes mitochondrial glutamine metabolism through HIF1α-GDH pathway in human lung cancer cells.

Drug-resistance is common in human lung cancer therapy. Hypoxia remarkably contributes to drug-resistance in lung cancer but the underlying mechanism remains elusive. Here we demonstrate that hypoxia-induced glutamine metabolism is involved in drug resistance in lung cancer cells. Hypoxia increases glutamine up-take, glutamate to α-ketoglutarate flux and the generation of ATP in lung cancer cells by up-regulating the expression of glutamate dehydrogenase (GDH). Hypoxia-induced expression of GDH relies on the up-regulation of HIF1α but not HIF2α. HIF1α binds the promoter of GDH and promotes the transcription of GDH gene in lung cancer cells. Finally, we show that GDH represses cisplatin-induced cell apoptosis and repression of colony formation, indicating that GDH contributes to drug-resistance in lung cancer cells. In conclusion, HIF1α-GDH pathway regulates glutamine metabolism and ATP production upon hypoxia stress and contributes to drug-resistance in human lung cancer cells.

Gemcitabine-Induced Autophagy Protects Human Lung Cancer Cells from Apoptotic Death.

PURPOSE: Gemcitabine has been used as a therapeutic drug combined with cisplatin for the treatment of lung cancer patients. However, the prognosis is poor due to acquired resistance. Accumulating studies have revealed that autophagy may contribute to the drug resistance. Therefore, the present study is aimed to clarify the mechanisms underlying gemcitabine-acquired resistance. METHODS: SPC-A1 and A549 cells were incubated with gemcitabine followed by assessment of cell viability with MTT assays. GFP-LC3 transient transfection, MDC staining, and transmission electron microscopy were used to detect the change of autophagy at morphological level. Flow cytometry was used to monitor the effect of 3-MA on gemcitabine-induced apoptosis. Western blot analysis was used to detect the expression of p62, LC3, Beclin-1, ATG5, activated caspase 3, Bax, BNIP3, BNIP3L, and Bcl-2. RESULTS: Our study showed that gemcitabine significantly induced both autophagy and apoptosis in human lung cancer cells SPC-A1 and A549. Of interest was that when autophagy was inhibited by 3-MA, the gemcitabine-induced apoptosis was effectively enhanced, suggesting that gemcitabine can activate autophagy to impair the chemosensitivity of lung cancer cells. Furthermore, the inhibition of autophagy by 3-MA further increased the expression of activated caspase 3, Bax, BNIP3, and BNIP3L, all are critical apoptotic mediators. Contrarily, 3-MA treatment further decreased the expression of Bcl-2, which is an important anti-apoptotic protein. CONCLUSION: Our study indicated that autophagy protected human lung cancer cells from gemcitabine-induced apoptosis, and the combined use of gemcitabine and an autophagic inhibitor in lung cancer patients may be an effective therapeutic strategy.

Poly-L-Arginine Acts Synergistically with LPS to Promote the Release of IL-6 and IL-8 via p38/ERK Signaling Pathways in NCI-H292 Cells.

Major basic protein (MBP) derived from activated eosinophil can exacerbate atopic asthma induced by lipopolysaccharide (LPS). The pharmacological function of MBP can be mimicked by poly-L-arginine (PLA), however, the potential signaling mechanisms of LPS-PLA-induced release of the inflammatory cytokines interleukin (IL)-6 and IL-8 remain unclear. In the present study, airway epithelia NCI-H292 cell lines were treated with LPS and/or PLA. We found that the expression levels of IL-6 and IL-8 induced by LPS-PLA were increased significantly compared with that in untreated cells. Meanwhile, the phosphorylation of p38 MAPK and ERK1/2 was also up-regulated dramatically by LPS-PLA, but this increase could be blocked by specific inhibitor. Importantly, blocking the phosphorylation of p38 MAPK and ERK1/2 reduced the expression levels of IL-6 and IL-8 as well. Collectively, LPS-PLA-induced release of IL-6 and IL-8 from NCI-H292 cells may be due to the synergistic activation of p38 MAPK and ERK1/2 signaling transduction pathways.

Amplicon Indel Hunter Is a Novel Bioinformatics Tool to Detect Large Somatic Insertion/Deletion Mutations in Amplicon-Based Next-Generation Sequencing Data.

Amplicon-based targeted next-generation sequencing assays are used widely to test for clinically relevant somatic mutations in cancer. However, accurate detection of large insertions and deletions (indels) via these assays remains challenging. Sequencing reads that cover these anomalies are, by definition, different from the reference sequence, and lead to variable performance of alignment algorithms. Reads with large indels may be aligned incorrectly, causing incorrect calls, or may remain unmapped and essentially ignored by downstream informatics pipeline sub-processes. Herein, we describe Amplicon Indel Hunter (AIH), a novel large (>5-bp) indel detection method that is reference genome independent and highly sensitive for the identification of somatic indels in amplicon-based, paired-end, next-generation sequencing data. We validated the algorithm for sensitivity and specificity using a set of clinical cancer samples with Clinical Laboratory Improvement Amendment-confirmed indels as well as in silico mutated data sets. The AIH detected 100% of tested large indels with relatively higher mutant allele frequencies compared with a variety of traditional aligners, which showed variably reduced sensitivity and specificity by comparison. The AIH especially outperformed alignment-based methods for detection of all tested FLT3 internal tandem duplications up to 102 bp. Because AIH runs in parallel to traditional alignment-based informatics pathways, it can be readily incorporated into nearly any analysis pipeline for somatic mutation detection in paired-end amplicon-based data.

Hypoxia-induced autophagy mediates cisplatin resistance in lung cancer cells.

Hypoxia which commonly exists in solid tumors, leads to cancer cells chemoresistance via provoking adaptive responses including autophagy. Therefore, we sought to evaluate the role of autophagy and hypoxia as well as the underlying mechanism in the cisplatin resistance of lung cancer cells. Our study demonstrated that hypoxia significantly protected A549 and SPC-A1 cells from cisplatin-induced cell death in a Hif-1α- and Hif-2α-dependent manner. Moreover, compared with normoxia, cisplatin-induced apoptosis under hypoxia was markedly reduced. However, when autophagy was inhibited by 3-MA or siRNA targeted ATG5, this reduction was effectively attenuated, which means autophagy mediates cisplatin resisitance under hypoxia. In parallel, we showed that hypoxia robustly augmented cisplatin-induced autophagy activation, accompanying by suppressing cisplatin-induced BNIP3 death pathways, which was due to the more efficient autophagic process under hypoxia. Consequently, we proposed that autophagy was a protective mechanism after cisplatin incubation under both normoxia and hypoxia. However, under normoxia, autophagy activation 'was unable to counteract the stress induced by cisplatin, therefore resulting in cell death, whereas under hypoxia, autophagy induction was augmented that solved the cisplatin-induced stress, allowing the cells to survival. In conclusion, augmented induction of autophagy by hypoxia decreased lung cancer cells susceptibility to cisplatin-induced apoptosis.

Reversed expression of GRIM-1 and GRP78 in human non-small cell lung cancer.

Gene associated with retinoid and interferon-induced mortality 1 (GRIM-1) acts as a tumor growth suppressor via apoptosis induction. However, GRIM-1 expression in human non-small cell lung cancer (NSCLC) and its potential interaction with another apoptosis-associated protein-glucose-regulated protein 78 (GRP78)-are as yet unknown. Using 40 surgical specimens, we showed significantly lower expression of GRIM-1 in NSCLC at both protein and messenger RNA (mRNA) levels compared with that in normal tissues (P < .01 and P < .001, respectively). Interestingly, these tumors tended to express higher basal amounts of GRP78 protein and mRNA (P < .05 and P < .001, respectively). Similarly, in the NSCLC tissues, weaker staining for GRIM-1 (main intensity + to ++) but stronger staining for GRP78 (main intensity +++ to ++++) was observed. Correlation analysis showed that protein and mRNA expression or the percentage of cells immunoreactive for GRIM-1 was negatively correlated with that of GRP78 (r = -0.279, r = -0.326, or r = -0.571, respectively). Coimmunoprecipitation and transient transfection revealed that GRIM-1 interacted with GRP78 and suppressed GRP78 protein expression. In addition, there was no correlation between GRIM-1 expression and clinical characteristics, whereas GRP78 expression was significantly correlated with tumor-nodes-metastasis (TNM) stage (stage 3 + 4 versus stage 1 + 2). In conclusion, the expression of GRIM-1 and GRP78 was negatively correlated in human NSCLC tissues, and the down-regulation of GRP78 by GRIM-1 provides a possible mechanism for their interaction. This study suggests a novel potential molecular pathway inactivated during the development of NSCLC.

Host sex-specific parasites in a functionally dioecious fig: a preference way of adaptation to their hosts.

Host-parasites interaction is a common phenomenon in nature. Diffusive coevolution might maintain stable cooperation in a fig-fig wasps system, in which the exploiter might diversify their genotype, phenotype, or behavior as a result of competition with pollinator, whereas the figs change flower syconia, fruits thickness, and syconia structure. In functionally dioecious Ficus auriculata, male figs and female figs contain two types of florets on separate plant, and share high similarities in outside morphology. Apocryptophagus (Sycophaginae, Chalcidoidea, Hymenoptera) is one of few groups of nonpollinating fig wasps that can reproduce within both male and female figs. On the basis of the morphology and DNA barcoding, evidence from partial sequences of mitochondrial cytochrome c oxidase I and nuclear internal transcribed spacer 2, we found that there are two nonsibling Apocryptophagus species living on male and female F. auriculata figs, respectively. We estimated that these two species diverged about 19.2 million years ago. Our study suggests that the host shift from Ficus variegate or Ficus prostrata fig species to male figs is a preference way for Apocryptophagus wasps to adapt to the separation of sexual function in diecious figs. Furthermore, to escape the disadvantage or sanction impact of the host, the exploiter Apocryptophagus wasps can preferably adapt to exploiting each sex of the figs, by changing their oviposition, niche shift, and habitat.

Pollinating fig wasp Ceratosolen solmsi adjusts the offspring sex ratio to other foundresses.

Local mate competition theory predicts that offspring sex ratio in pollinating fig wasps is female-biased when there is only one foundress, and increased foundress density results in increased offspring sex ratio. Information of other foundresses and clutch size have been suggested to be the main proximate explanations for sex ratio adjustment under local mate competition. Our focus was to show the mechanism of sex ratio adjustment in a pollinating fig wasp, Ceratosolen solmsi Mayr, an obligate pollinator of the functionally dioecious fig, Ficus hispida Linn., with controlled experiments in the field. First, we obtained offspring from one pollinator and offspring at different oviposition sequences, and found that offspring sex ratio decreased with clutch size, and pollinators produced most of their male offspring at the start of bouts, followed by mostly females. Second, we found that offspring sex ratio increased with foundress density, and pollinators did adjust their offspring sex ratio to other females in the oviposition patches. We suggest that when oviposition sites are not limited, pollinators will mainly adjust their offspring sex ratio to other foundresses independent of clutch size changes, whereas adjusting clutch size may be used to adjust sex ratio when oviposition sites are limited.

Comparison of the genome sequences of "Candidatus Portiera aleyrodidarum" primary endosymbionts of the whitefly Bemisia tabaci B and Q biotypes.

"Candidatus Portiera aleyrodidarum" is the primary endosymbiont of whiteflies. We report two complete genome sequences of this bacterium from the worldwide invasive B and Q biotypes of the whitefly Bemisia tabaci. Differences in the two genome sequences may add insights into the complex differences in the biology of both biotypes.

Genome sequences of the primary endosymbiont "Candidatus Portiera aleyrodidarum" in the whitefly Bemisia tabaci B and Q biotypes.

"Candidatus Portiera aleyrodidarum" is the obligate primary endosymbiotic bacterium of whiteflies, including the sweet potato whitefly Bemisia tabaci, and provides essential nutrients to its host. Here we report two complete genome sequences of this bacterium from the B and Q biotypes of B. tabaci.

Expression and clinical significance of GRIM-19 in lung cancer.

We investigated the expression of gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) in lung cancer, a recently discovered cell death regulatory gene. Over-expression of GRIM-19 potentially suppresses proliferation and promotes tumor cell apoptosis. However, the expression of GRIM-19 in human lung cancer has not yet been thoroughly investigated. All of the specimens were obtained using CT-guided lung puncture or bronchial biopsy. The expression of GRIM-19 was investigated using immunohistochemistry. The expression level of GRIM-19 was significantly different between lung cancer and lung inflammation. A relatively lower GRIM-19 expression level was also found in small cell lung carcinomas compared to squamous cell carcinoma and adenocarcinoma. No significant difference between GRIM-19 expression in squamous cell carcinoma and adenocarcinoma was determined. Downregulation of GRIM-19 was found in non-small cell lung carcinomas stages III-IV compared to stages I-II, indicating a negative correlation between the expression level of GRIM-19 and the stage of the primary lesion (T). Furthermore, we found GRIM-19 to be primarily located in the cytoplasm in lung inflammation tissues, but located in the nucleus in lung cancer tissues. GRIM-19 expression occurs as an early phenomenon in the pathogenesis of lung cancer. Our study found that GRIM-19 expression in lung cancer is significantly lower compared to lung inflammation, exhibits a relationship with the histological type and clinical stage of lung cancer, and is a suitable target for the development of new lung cancer therapies.

Decreased expression of Beclin-1 and LC3 in human lung cancer.

Autophagy, a conversed response to stress, has recently been studied in human cancers. Two important autophagic genes-Beclin-1 and LC3 are reported in several human cancers. However, the expressions of Beclin-1 and LC3 in lung cancer have not yet been investigated. In the present study, we investigated the expression of Beclin-1 and LC3, and the relationship between the expression profile and the clinical or pathological changes in human lung cancer. 40 primary lung cancer patients are involved in present study. mRNA expressions of Beclin-1 and LC3-II were detected by Real Time PCR and the protein levels were assessed by immunohistochemistry and western blot. Relative lower expressions of Beclin-1 and LC3-II mRNA were found in the lung cancer tissues compared to counterpart normal tissues. Consistently, the lower amount of Beclin-1 and LC3-II protein was found in lung cancer tissues. However, the expressions of Beclin-1 and LC3-II in lung cancer tissues were not affected by patients' age, gender, smoking, histological type, lymph node metastasis and tumor-node-metastasis (TNM) stage. Both mRNA and protein levels of Beclin-1 and LC3-II were significantly decreased in lung cancer tissues which suggested that autophagy may be involved in the pathogenesis of lung cancer.

Gene associated with retinoid-interferon-induced mortality-19 suppresses growth of lung adenocarcinoma tumor in vitro and in vivo.

Lung cancer is currently the leading cause of cancer-related death in the world. Signal transducers and activators of transcription 3 (STAT3) is a major oncogenic transcription factor involved in the development and progression of a number of human tumors including lung denocarcinoma. Gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) is known to functionally interact with STAT3 and inhibit its transcriptional activity. Decreased expression of GRIM-19 has been reported in tumors including those from kidney, prostate, colon and cervix, indicating that loss of GRIM-19 may be involved in the tumorigenesis through activation of the STAT3 pathway. In this study, we determined that GRIM-19 was significantly reduced at the mRNA and protein levels in lung adenocarcinoma tissues. Moreover, STAT3 was increased in these tumors and corresponding changes in the expression of its downstream target genes was observed. Overexpression of GRIM-19 was also found to suppress lung adenocancinoma tumor growth both in vitro and in vivo. Taken together, these findings will likely contribute to the future development of GRIM-19-based gene therapy approaches to treat lung adenocancinoma.

Enrichment of mRNA-like noncoding RNAs in the divergence of Drosophila males.

With the advent of transcriptome data, it has become clear that mRNA-like noncoding RNAs (mlncRNAs) are widespread in eukaryotes. Although their functions are poorly understood, these transcripts may play an important role in development and could thus be involved in determining developmental complexity and phenotypic diversification. However, few studies have assessed their potential roles in the divergence of closely related species. Here, we identify and study patterns of sequence and expression divergence in ten novel candidate mlncRNAs from Drosophila pseudoobscura and its close relative D. persimilis. The candidate mlncRNAs were identified by randomly sequencing a group of 734 cDNA clones from a microarray that showed either no difference in expression (187 clones) or differential expression (547 clones) in comparisons between D. pseudoobscura and D. persimilis and between these two species and their F(1) hybrids. Candidate mlncRNAs are overrepresented among differentially expressed transcripts between males of D. pseudoobscura and D. persimilis, and although they have high sequence conservation between these two species, seven of them have no putative homologs in any of the other ten Drosophila species whose genomes have been sequenced. Expression of eight of the ten candidate mlncRNAs was detected either in whole bodies (adults) or testes using a custom-designed oligonucleotide microarray. Three of the ten candidate mlncRNAs are highly expressed (in the top 4% of the male transcriptome), differentially expressed between species, and show extreme levels of sex-bias, with one transcript having the highest level of male bias in the whole transcriptome. Proteomic data from testes show no traces of any predicted peptides from the candidate mlncRNAs. Our results suggest that these mlncRNAs may be important in male-specific processes related to sexual dimorphism and species divergence in this species group.

Evolution of sex-dependent gene expression in three recently diverged species of Drosophila.

Sexual dimorphism in morphological, physiological, and behavioral traits is pervasive in animals, as is the observation of strong sexual dimorphism in genomewide patterns of gene expression in the few species where this has been studied. Studies of transcriptome divergence show that most interspecific transcriptional divergence is highly sex dependent, an observation consistent with the action of sex-dependent natural selection during species divergence. However, few transcriptome evolution studies have been conducted between recently diverged species (<1 MY). Here, we present analyses of sex-biased transcriptome divergence in sexually mature adults of three recently diverged species of Drosophila: Drosophila pseudoobscura, D. persimilis, and D. pseudoobscura bogotana. Data were collected using a custom designed Agilent oligonucleotide. Expression was detected in 12,507 genes. About 80% of the expressed genes show sex-biased expression in each species. Across species, 21% of the transcriptome shows switches between nonsex bias and sex bias, and just 0.9% of the transcriptome shows reversals of sex-biased expression. Over 80% of the expression divergence between species is due to changes in one sex only. About 15% of the expression divergence between species is due to changes in the same direction in both sexes and just 2% is due to changes in both sexes but in opposite directions. In agreement with previous studies, we observe a high level of sex-dependent transcriptome divergence and strong demasculinization of the two arms of the X chromosome in all species. However, in contrast to previous studies we find that male-biased genes do not have higher levels of expression divergence than non-sex-biased genes, and sex-biased genes show higher levels of expression divergence in the alternate sex, suggesting that sex-biased genes endure stronger selection when expressed in the alternate sex.

New insights into the phylogeny of fig pollinators using Bayesian analyses.

The interaction between figs and fig pollinators is one of the most species-specific mutualisms. Recently, phylogenies of both partners based on molecular data provided insights into a wide spectrum of co-evolutionary questions. However, for the phylogeny of fig pollinators, there are some discrepancies between different studies and left some relationships unresolved, especially for deep nodes. The phylogenetic uncertainties of pollinators prohibit our further understanding of the history of the mutualism. Here, we present phylogenetic analyses of a larger COI sequence dataset that includes previously published datasets and our sequences from 20 species using Bayesian method and maximum parsimony. The analyses using different methods share similar topologies. Bayesian analyses provide high level of confidence for most internal nodes in terms of posterior probability. This study also clarifies some discrepancies between previous studies. After rooting with Tetrapus, other pollinators split into two clades. Wiebesia and Blastophaga are at basal positions in respective clade. Ceratosolen is not monophyletic because Kradibia and Liporrhopalum fall inside this group. Three subgenera of Ceratosolen: subgen. Ceratosolen, subgen. Rothropus, and subgen. Strepitus are not supported. Therefore, Ceratosolen is suggested to be re-divided into three groups. Urostigma pollinators (including Dolichoris and Blastophaga psenes) are clustered together. The monophylies of Wiebesia, Blastophaga, Dolichoris are not supported in this analysis. This study also provides a new framework for re-evaluating character evolution and re-inspecting the definition of some genera.