Abstract Patterning and phenotypic variations of appendages in insects provide important clues on developmental genetics. In the silkworm B ombyx mori , morphological variations associated with the E complex, an analogue of the D rosophila melanogaster bithorax complex, mainly determine the shape and number of prolegs on abdominal segments. Here, we report the identification and characterization of the allele responsible for the supernumerary crescents and legs‐like ( E Cs ‐l ) mutant, a model derived from spontaneous mutation of the E complex, with supernumerary legs and extra crescents. Fine mapping with 1605 individuals revealed a ∼68 kb sequence in the upstream intergenic region of B . mori abdominal‐ A ( B mabd‐ A ) clustered with the E Cs ‐l locus. Quantitative real‐time PCR (q RT‐PCR ) and W estern blotting analyses disclosed a marked increase in B mabd‐ A expression in the E Cs ‐l mutant at both the transcriptional and translational levels, compared to wild‐type D azao . Furthermore, we observed ectopic expression of the Bmabd‐ A protein in the second abdominal segment ( A 2) of the E Cs ‐l mutant. Our results collectively suggest that the 68 kb region contains important regulatory elements of the B mabd‐ A gene, and provide evidence that the gene is required for limb development in abdominal segments in the silkworm.
Flight loss has occurred in many winged insect taxa. The flightless silkmoth Bombyx mori, is domesticated from the wild silkmoth, Bombyx mandarina, which can fly. In this paper, we studied morphological characteristics attributed to flightlessness in silkmoths. Three domestic flightless B. mori strains and one B. mandarina population were used to compare morphological components of the flight apparatus, including wing characteristics (shape, forewing area, loading, and stiffness), flight muscle (weight, ratio, and microscopic detail) and body mass. Compared with B. mandarina, B. mori strains have a larger body, greater wing loading, more flexible wings and a lower flight muscle ratio. The arrangement in microscopy of dorsal longitudinal flight muscles (DLFMs) of B. mori was irregular. Comparative analysis of the sexes suggests that degeneration of flight muscles and reduction of wing mechanical properties (stiffness) are associated with silkmoth flightlessness. The findings provide important clues for further research of the molecular mechanisms of B. mori flight loss.
Human sepiapterin reductase (SR) deficiency is an inherited disease caused by SPR gene mutations and is a monoamine neurotransmitter disorder. Here, we investigated whether the silkworm lemon mutant could serve as a model of SR deficiency. A point mutation in the BmSPR gene led to a five amino acid deletion at the carboxyl terminus in the lemon mutant. In addition, classical phenotypes seen in SR deficient patients were observed in the lemon mutant, including a normal phenylalanine level, a decreased dopamine and serotonin content, and an increased neopterin level. A recovery test showed that the replenishment of l -dopa significantly increased the dopamine level in the lemon mutant. The silkworm lemon mutant also showed negative behavioural abilities. These results suggest that the silkworm lemon mutant has an appropriate genetic basis and meets the biochemical requirements to be a model of SR deficiency. Thus, the silkworm lemon mutant can serve as a candidate animal model of SR deficiency, which may be helpful in facilitating accurate diagnosis and effective treatment options of SR deficiency.
The silkworm neuropeptide Orcokinin (abbreviated as BommoOK) is equipped with multiple biological functions, one of which acts as a pigmentation inhibitor. To explore the whitening efficiency of BommoOK, the inhibitory effects on tyrosinase and its adaptability on the cell for six mature peptides of BommoOK were investigated in this paper. At the same time, BommoOKA_type4, the peptide with the best melanin inhibition effect, was used as an additive to prepare a whitening cream, and the effects on skin moisture, oil content, fine lines, skin glossiness, pores, and pigment depth were determined. The results revealed that the cream added with BommoOKA_type4 peptide showed a good improvement effect on the skin, especially in significantly reducing the pigment depths of skin. The results displayed a potential application of BommoOK in whitening and skincare products as an excellent additive and provide certain references for the mechanism research of BommoOK in inhibiting melanin synthesis.
Cirrhotic patients with hemorrhagic ascites have significant morbidity and mortality. This study aims to determine the relationship between D-dimer values and hemorrhagic ascites in cirrhotic patients and analyze its predictive value.This retrospective study screened 572 consecutive cirrhotic patients with ascites and hemorrhagic ascites (defined as red blood cells (RBC) in ascitic fluid ≥ 10,000/µL) during a 72-month period. The overall patient survival rate was measured by Kaplan-Meier analysis method. The relationship between D-dimer and hemorrhagic ascites was also examined. A multivariate Cox proportional hazard analysis was performed to assess the indepen-dent risk factors related to mortality.Both control group and hemorrhagic ascites patients had obvious hepatic dysfunction as determined by Model for End-Stage Liver Disease (MELD) scores of 6.37 ± 1.05 and 11.82 ± 2.86, respectively (p < 0.001). There was a higher prevalence of patients with significant ascites in those with spontaneous hemorrhagic ascites than in the control group (p = 0.003). There were significant differences in D-dimer levels between both groups (9.44 ± 5.11 vs. 26.83 ± 5.35, p < 0.001). Hemorrhagic ascites was significantly and positively correlated with D-dimer levels (r = 0.692, p < 0.0001). The area under the receiver operating characteristic (ROC) curve was 0.9838. Using Cox proportional hazard model for multivariate prognostic analysis, MELD, D-dimer and presence of spontaneous hemorrhagic ascites were independent predictors of 3-year mortality.Patients with hemorrhagic ascites had a significantly higher MELD score, D-dimer, and mortality than patients with ascites alone. D-dimer was associated with the appearance of hemorrhagic ascites and was found to be a marker of advanced liver disease and poor outcomes.
Cytosine arabinoside (Ara-c) is a pyrimidine anti-metabolite that is capable of interfering with cellular proliferation by inhibiting DNA synthesis. Each inhibitor of cyclin-dependent kinase 4 (INK4) family member has the ability to bind to cyclin-dependent kinase 4 (CDK4) and inhibit the formation of the cell cycle-dependent CDK4/cyclin D1 complex, subsequently leading to cell cycle arrest in the G1/S phase. In this study, the expression of INK4 family genes in kidney cancer and the impact of these genes on patient prognosis were examined. Additionally, the effects of INK4 family genes and Ara-c on cell proliferation and tumor formation and development were examined. Finally, a potential association between Ara-c-induced cell cycle arrest and INK4-associated gene expression was evaluated. An upregulation of INK4 family genes was found to be positively correlated with the prognosis of patients with kidney cancer. Both the INK4 family genes and Ara-c were shown to induce cell cycle arrest and inhibit tumor formation and development. Moreover, Ara-c-induced cell cycle arrest was found to be associated with an Ara-c-induced upregulation of INK4 family gene expression, which ultimately inhibited the formation of the CDK4/cyclin D1 complex. These findings suggested that an upregulation of INK4 family genes has a positive effect on kidney cancer prognosis and can inhibit the formation and development of tumors. Moreover, Ara-c was shown to promote the upregulation of INK4 family genes, at the same time, Ara-c could directly regulate the cell cycle-dependent genes CDK4 and cyclin D1 (CCND1), independent of the INK4 family genes.
Introduction: SHR0302 is a highly selective JAK1 inhibitor. This study aims to investigate the safety, tolerability, and pharmacokinetics of single- and multiple-dose topical skin application of SHR0302 base ointment in healthy adult subjects. Methods: This phase I clinical trial (registration number: CTR20192188) consisted of two parts. Part 1 was a single-dose ascending study with four dose levels in 32 healthy Australian adults (eight subjects in each dose group). All Australian subjects were randomized 3:1 to a single-dose topical skin application of SHR0302 base ointment or placebo. The dose escalated from 1% SHR0302 base ointment on 3% of body surface area (BSA) to 2% SHR0302 base ointment on 20% of BSA. Part 2 combined single- and multiple-dose-ascension studies with two dose levels in 20 healthy Chinese adults (10 subjects in each dose group). All Chinese subjects were randomized 4:1 to a combination of single and multiple doses for consecutive 10 days of topical application of 1% SHR0302 base ointment on 20% BSA or 2% SHR0302 base ointment on 20% BSA. The safety and pharmacokinetics of the SHR0302 base ointment were evaluated. Results: The incidence of treatment-emergent adverse events (TEAEs) in both parts was comparable between the SHR0302 base ointment group and the vehicle group (Part 1:33.3% vs. 37.5%; Part 2: 56.3% vs. 75.0%). All TEAEs were transient, recovered, and equally well-tolerated in the two racial groups. The overall absorption of the SHR0302 base ointment was slow after topical application, with Tmax>10 h. After a single dose of the SHR0302 base ointment, drug exposure in healthy Australian and Chinese subjects increased non-linearly with the increase in the administration area and drug content. Drug exposure increased in a less-than-dose-proportional manner within the dose range tested. Due to differences in the clinical practice of topical application, the Tmax of the drug in Australian subjects was earlier than in Chinese subjects, but the overall extent of absorption seemed comparable in Australian and Chinese subjects (with comparable AUC0-t). Conclusion: The SHR0302 base ointment (either single or multiple doses) was well tolerated and safe, with no racial disparity.
The color pattern of insects is one of the most dazzling adaptive evolutionary phenotypes. However, the molecular regulation of this color pattern is not fully understood. In this study, we found that the transcription factor Bm-mamo is responsible for black dilute ( bd ) allele mutations in the silkworm. Bm-mamo belongs to the BTB zinc finger family and is orthologous to mamo in Drosophila melanogaster . This gene has a conserved function in gamete production in Drosophila and silkworms and evolved a pleiotropic function in the regulation of color patterns in caterpillars. We found that Bm-mamo can comprehensively regulate the expression of related pigment synthesis and cuticular protein genes to form color patterns. This indicates that insects have a genetic basis for coordinate regulation of the structure and shape of the cuticle, as well as color patterns. This genetic basis provides the possibility for constructing the complex appearances of some insects. This study provides new insight into the regulation of color patterns.The color patterns of insects are highly exquisite and significantly divergent. The metabolism of pigments is the material basis for insect coloration. However, the cuticle of insects plays an important role as a scaffold for carrying pigment particles. Cuticular proteins are some of the main components of the cuticle. This study showed that a BTB-ZF family transcription factor protein, Bm-mamo, can comprehensively regulate melanin synthesis and the expression of multiple cuticular protein genes. Hence, insects have a genetic basis for integrated control of cuticle and color pattern construction, which enables them to produce complex appearances.
// Jiangbo Song 1,2,* , Dongmei Tang 1,2,* , Zhiquan Li 1,2,* , Xiaoling Tong 1,2 , Jianfei Zhang 1,2 , Minjin Han 1,2 , Hai Hu 1,2 , Cheng Lu 1,2 and Fangyin Dai 1,2 1 State Key Laboratory of Silkworm Genome Biology, College of Biotechnology, Southwest University, Chongqing, China 2 Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, College of Biotechnology, Southwest University, Chongqing, China * These authors are co-first authors Correspondence to: Fangyin Dai, email: // Keywords : silkworm, lifespan, dietary restriction, BmFoxO, experimental animal, Gerotarget Received : September 27, 2016 Accepted : December 20, 2016 Published : December 26, 2016 Abstract Established animal models have accelerated our understanding of the mechanisms involved in lifespan determination. However, more experimental animals are required to clarify the complex mechanisms behind the phenomena of aging and lifespan. In this study, we reported the variation of lifespan in nine distinct silkworm strains. Lifespan correlated significantly with BmFoxO gene expression in the representative silkworm strains tested (Xiafang, Dazao-N, and N4). In general, the female silkworm was longer lived than the male of the same strain. Dietary restriction extended the silkworm lifespan compared with that of silkworms fed ad libitum . The expression of BmFoxO was significantly elevated in the dietary restriction group on day 3 of the 4 th instar and day 3 of the 5 th instar, suggesting that BmFoxO contributes to dietary restriction-mediated lifespan extension. The RNA interference and overexpression of the BmFoxO gene significantly shortened and extended the silkworm adulthood, respectively. In conclusion, our findings suggest that the silkworm might serve as a promising experimental animal to explore the complex biological mechanisms of lifespan determination.
Abstract Long noncoding RNAs (lncRNAs) areinvolvedin a variety of biological processes. In silkworm, numerous lncRNAs have been predicted through deep transcriptome sequencing, but no functional role has been experimentally validated yet. Here, we characterized a new lncRNA iab‐1 that was mainly encoded by the intergenic region between Bmabd‐A and Bmabd‐B in the Homeobox (Hox) cluster of the silkworm, Bombyx mori . More than seven alternative splicing isoforms of lncRNA iab‐1 were cloned, which were subgrouped into types 1 and 2 based on the location of the 3′‐ends. The iab‐1 was expressed at a low level, but the expression of iab‐1 peaked at several specific development stages, including 3 to 4 days during the embryonic stage, stages before fourth molting, and the sixth hour after the fourth molting, and early stages during metamorphosis. It was highly expressed in the nervus and epidermis, especially the epidermis of the posterior abdomen at the fourth instar premolting stage. The relationship between iab‐1 and nearby Hox genes was analyzed at different developmental stages. Iab‐1 expression was highly associated with Bmabd‐A as well as Bmabd‐B in the embryonic and larval stages, while this association was decreased at the metamorphic stage; iab‐1 expression was highly associated with BmUbx only in the embryonic stage. Downregulation of iab‐1 expression by small interfering RNA led to the death of most of the treated individuals at the larval stage, suggesting that iab‐1 transcript expression might be involved in certain relevant physiological processes. The expression of Bmabd‐A and Bmabd‐B did not change in iab‐1 downregulated individuals, indicating that the relevance between the two genes and iab‐1 was not induced by iab‐1 transcript. Collectively, the results showed that the newly identified iab‐1 may be involved in some physiological processes, and the interaction between iab‐1 and Hox genes was also preliminarily analyzed.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)