Bombyx mori RPL13 participates in UV-induced DNA damage repair of B. mori nucleopolyhedrovirus through interaction with Bm65.

Ribosomal protein L13 (RPL13) is highly conserved in evolution. At present, the properties and functions of RPL13 have not been characterised in insects. In this study, Bombyx mori RPL13 (BmRPL13) was first found to be specifically recruited to the sites of ultraviolet (UV)-induced DNA damage and contributed to UV damage repair. Escherichia coli expressing BmRPL13 showed better resistance to UV radiation. After knocking down the expression of BmRPL13 in BmN cells, the repair speed of UV-damaged DNA slowed down. The further results showed that BmRPL13 interacted with B. mori nucleopolyhedrovirus (BmNPV) ORF65 (Bm65) protein to locate at the UV-induced DNA damage sites of BmNPV and helped repair UV-damaged viral DNA.

Characterization of the UDP-glycosyltransferase UGT33D1 in silkworm Bombyx mori.

Uridine diphosphate (UDP)-glycosyltransferases (UGTs) are important metabolizing enzymes functioning by adding a sugar moiety to a small lipophilic substrate molecule and play critical roles in drug/toxin metabolism for all realms of life. In this study, the silkworm Bombyx mori UGT33D1 gene was characterized in detail. UGT33D1 was found localized in the endoplasmic reticulum (ER) compartment just like other animal UGTs and was mainly expressed in the silkworm midgut. We first reported that UGT33D1 was important to BmNPV infection, as silencing UGT33D1 inhibited the BmNPV infection in silkworm BmN cells, while overexpressing the gene promoted viral infection. The molecular pathways regulated by UGT33D1 were analysed via transcriptome sequencing upon UGT33D1 knockdown, highlighting the important role of the gene in maintaining a balanced oxidoreductive state of the organism. In addition, proteins that physically interact with UGT33D1 were identified through immunoprecipitation and mass spectrometry analysis, which includes tubulin, elongation factor, certain ribosomal proteins, histone proteins and zinc finger proteins that had been previously reported for human UGT-interacting proteins. This study provided preliminary but important functional information on UGT33D1 and is hoped to trigger deeper investigations into silkworm UGTs and their functional mechanisms.

BmNPV Orf 65 (Bm65) Is Identified as an Endonuclease Directly Facilitating UV-Induced DNA Damage Repair.

Baculoviruses have been used as biopesticides for the control of Lepidoptera larvae. However, solar UV radiation reduces the activity of baculovirus. In this study, an UV endonuclease, Bm65, was found encoded in the genome of Bombyx mori nuclear polyhedrosis virus (BmNPV). Bm65 (the ortholog of AcMNPV orf79) was guided by a key nuclear localization signal to enter the nucleus and accumulated at UV-induced DNA damage sites. Subsequent results further showed that Bm65-mediated DNA damage repair was not the only UV damage repair pathway of BmNPV. BmNPV also used host DNA repair proteins to repair UV-induced DNA damage. In summary, these results revealed that Bm65 was very important in UV-induced DNA damage repair of BmNPV, and BmNPV repaired UV-damaged DNA through a variety of ways. IMPORTANCE Baculovirus biopesticides are environmentally friendly insecticides and specifically infect invertebrates. UV radiation from the sunlight greatly reduces the activity of baculovirus biopesticides. However, the molecular mechanisms of most baculoviruses to repair UV-induced DNA damage remain unclear. Nucleotide excision repair (NER) is a major DNA repair pathway that removes UV-induced DNA lesions. At present, there are few reports about the nucleotide excision repair pathway in viruses. Here, we showed for the first time that the baculovirus Bm65 endonuclease actually cleaved UV-damaged DNA. Meanwhile, we found that BmNPV used both viral-encoded enzymes and host DNA damage repair proteins to reverse UV-induced DNA damage. These results will provide a reference for the research of UV damage repair of other viruses.

Research progress on nucleic acid detection and genome editing of CRISPR/Cas12 system.

PURPOSE: This work characterizes the applications of CRISPR/Cas12 system, including nucleic acid detection, animal, plant and microbial genome editing. METHODS: The literature on CRISPR/Cas12 system was collected and reviewed. RESULTS: CRISPR/Cas system is an acquired immune system derived from bacteria and archaea, which has become the most popular technology around the world because of its outstanding contribution in genome editing. Type V CRISPR/Cas systems are distinguished by a single RNA-guided RuvC nuclease domain with single effector molecule. Cas12a, the first reported type V CRISPR/Cas system, targets double-stranded DNA (dsDNA) adjacent to PAM sequences and trans-cleaves single-stranded DNA (ssDNA). We present the applications of CRISPR/Cas12 system for nucleic acid detection and genome editing in animals, plants and microorganisms. Furthermore, this review also summarizes the applications of other Cas12 proteins, such as Cas12b, Cas12c, Cas12d, and so on, which further widen the application prospects of CRISPR/Cas12 system. CONCLUSIONS: Knowledge of the applications of CRISPR/Cas12 system is necessary for improving the understanding of the functional diversity of CRISPR/Cas12 system and also provides significant references for further research and utilization of CRISPR/Cas12 in other new fields.

Effects of tolfenpyrad exposure on development and response mechanism in the silkworm, Bombyx mori.

Tolfenpyrad is a broad spectrum of insecticide that can effectively kill different types of pests, including Lepidoptera. However, due to improper use, the adverse effects of tolfenpyrad on beneficial or economic insects have not been well studied. In this study, we systematically investigated the toxic effect of sublethal tolfenpyrad on silkworms. Sublethal tolfenpyrad exposure can affect the body weight, developments days, cocooning rate, eclosion rate and pupation rate. To further study the response mechanism of silkworms to tolfenpyrad stimulation, we compared the different expression genes by transcriptome sequencing and verified them by qRT-PCR. We found that significant changes in the genes expression was involved in xenobiotics biodegradation and metabolism, immune system and digestive system after tolfenpyrad treatment. To further investigate the possible mechanisms by which intestinal microbia in the response to tolfenpyrad, we analysed the microbia changes in the midgut of silkworms by 16S rRNA gene sequencing. The results showed that the relative abundances of Enterobacter and Staphylococcus were increased whereas the Tyzzerella and Methylobacterium-Methylorubrum were decreased after tolfenpyrad stimulation. Taken together, these results indicated that low concentration of tolfenpyrad affect the growth and development of silkworms. Silkworms respond to the toxicity of tolfenpyrad by inducing immune and detoxification-related gene expression or altering microbial composition in the midgut.

Proteomic and Targeted Metabolomic Studies on a Silkworm Model of Parkinson's Disease.

Parkinson's disease (PD) is a chronic and progressive movement disorder that is characterized by the loss of dopaminergic neurons in the brain. Animal models of PD have become very popular in the past two decades to understand the etiology, pathology, and molecular and cellular pathways associated with PD. In this study, we report the first neurotoxin-induced silkworm model for PD by chronic feeding with 6-hydroxydopamine (6-OHDA) and explore the possible molecular mechanisms associated with PD using proteomic and targeted metabolomic approaches. Although silkworm is phylogenetically distant from humans and rats, 6-OHDA treatment produced similar PD phenotypes, including motor dysfunction, dopaminergic neuron degeneration, and decreased levels of dopamine. Major neurotransmitters in the silkworm head tissue were profiled, revealing key molecules implicating neurodegenerative disorder. Proteomics analysis revealed a major downregulation of nearly 50 structural proteins constituting cuticles and microfilaments, indicating mechanical damage in the silkworm tissues. The results suggest that 6-OHDA treatment could induce PD-like symptoms in silkworms and activate similar proteomic and metabolic pathways to those in rats or higher animals. This study demonstrates the feasibility and value of the silkworm-based PD model, which may provide important clues for understanding the molecular and cellular mechanisms underlying PD. The mass spectrometry raw files have been deposited to iProx via the project ID IPX0004206000.

An ultrasensitive electrochemical sensing platform based on silver nanoparticle-anchored 3D reduced graphene oxide for rifampicin detection.

A facile strategy has been reported to anchor silver nanoparticles (Ag NPs) onto three-dimensional reduced graphene oxide (3D rGO) via a green and simple method. An accurate and reliable electrochemical sensing platform based on Ag NPs/3D rGO was designed for the ultrasensitive detection of rifampicin (RIF). The morphology and features of Ag NPs/3D rGO were characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), Raman spectroscopy and electrochemical measurements. The interface of the modified electrode presented effective electrical activity for the analysis of RIF due to the large electrochemically active surface area and excellent electron transport ability. The sensor exhibited a good linear relationship in the range of 0.01 nM-45 µM and a low detection limit of 0.810 nM (S/N = 3). Crucially, the fabricated Ag NPs/3D rGO sensor was successfully utilized to assess RIF in human blood, drug and aquatic product samples. This sensing platform exhibited outstanding electrochemical performance for RIF detection and showed great potential application in clinical diagnosis, pharmaceutical and food-related fields.

Genomics and proteomics combined analysis revealed the toxicity response of silkworm Bombyx mori to the environmental pathogen Bacillus cereus ZJ-4.

Bacterial contamination has caused a major public health problem worldwide. Bacillus cereus is a conditional environmental pathogenic bacteria that can cause food poisoning. Whether environmental pathogens can cause widespread transmission in the insect kingdom is unclear. In this study, a Bacillus cereus ZJ-4 was isolated from the hospital environment of Zhenjiang City, Jiangsu Province, China. It was fatal by injection into the silkworm hemolymph. To investigated the potential toxic factors of ZJ-4 and clarified the toxicity response mechanism of silkworm by the ZJ-4 infection. Then, the whole genome of ZJ-4 was sequenced, and the immune mechanism of silkworm fat body to ZJ-4 pathogen was studied by HE pathological section and proteomics. Bacterial genome sequencing indicated that ZJ-4 had 352 drug resistance genes and 6 virulence genes. After 36 h of subcutaneous puncture with ZJ-4 suspension, the pathological changes were obviously found in HE pathological sections of fat body tissue. Comparative proteomic results indicated that differentially expressed proteins are mainly involved in stress reactions, biological regulation, and innate immunity. The qRT-PCR analysis showed that the expressions of ß-GRP, Spaetzle, MyD88, Tube and Dorsal genes in Toll pathway were up-regulated, while Pell and Cactus genes were down-regulated; in the antimicrobial peptide pathway, Glv2, Lzm, Mor, and Leb3 genes were up-regulated, while attacin1 and defensin genes were down-regulated; Sod gene was up-regulated, while Cat gene was down-regulated in the antioxidant pathway; Ldh, Sdh, and Mdh genes were down-regulated in glucose metabolism pathway. These results indicated that ZJ-4 can damage the innate immune pathway of silkworm, and also affect the normal immune function of fat body cells.

Identification, expression and function of myosin heavy chain family genes in Tribolium castaneum.

Functions of myosin heavy chain (myosin) family genes are poorly understood in some insects. To address this, we determined the expression and function of myosin family genes in Tribolium castaneum. TcMyo15 is predominantly expressed in early embryos, late larvae and early adults, but TcMyo7B transcripts significantly increased in late larvae. TcMyo20 transcripts are abundant in pre-adults, whereas TcMhc1 is highly expressed in post-embryonic stages. TcMhc2 shows peak expression in late pupae. TcMyo9 transcripts reached their highest levels in late pupae. TcMyo15, TcMyo7B and TcMhc2 are abundantly expressed in the adult epidermis, gut and testis, respectively. TcMyo9 and TcMyo20 are highly expressed in the epidermis, fat body and ovary, and TcMhc1 exhibits high mRNA levels in the epidermis and accessory gland. TcMyo20 RNAi reduced wing and leg size, fertility and egg hatchability. TcMyo9 knockdown completely inhibited eclosion and fecundity, but TcMyo15 or TcMhc1 silencing only impaired eclosion. TcMhc2 RNAi affected pupation and wing development. This study suggests myosin genes as potential targets of pesticides due to their critical roles in insect development and reproduction.

Silver nanoparticle toxicity in silkworms: Omics technologies for a mechanistic understanding.

The widespread use of silver nanoparticles (AgNPs) has raised public concern due to their potential toxic effects on humans and the environment. Although some studies have evaluated the toxicity of nanomaterials in vertebrates, studies on their hazardous effects on insects are limited. Here we focused on different concentrations of AgNPs to silkworms, a promising model organism, to evaluate their toxic effects by omics analysis. After the silkworms were fed with 100 mg L-1 AgNPs, transcriptomics analysis showed differential expression of 43 genes: 39 upregulated and 4 downregulated. These differentially expressed genes (DEGs) were involved in the digestion process, various metabolic pathways, transmembrane transport and energy synthesis. Proteomic results for silkworms fed with 400 mg L-1 AgNPs revealed 14 significantly differentially expressed proteins: 11 downregulated and 3 upregulated. Reverse transcription-polymerase chain reaction (RT-PCR) results showed that the expression levels of eight proteins were similar to the transcription levels of their corresponding genes. As the AgNPs concentration was increased, the expression of digestive enzymes was downregulated, which damaged the silkworm tissue and suppressed the activity of the enzyme superoxide dismutase and the protein HSP 1, causing oxidative stress and the production of reactive oxygen species, which had toxic effects on the silkworm digestive system. Histopathological results showed that treatment with 400 mg L-1 AgNPs destroyed the basal lamina and the columnar cells, caused adverse effects on tissues and had the potential to induce harmful effects on the digestive system. The data presented herein provide valuable information on the hazards and risks of nanoparticle contamination. Main finding: AgNPs would downregulate some digestive enzymes, damage the tissue of midgut in silkworm, meantime induce the accumulation of reactive oxygen species which may cause oxidative stress.

Crinkled employs wingless pathway for wing development in Tribolium castaneum.

Crinkled is associated with embryonic denticle formation and auditory organ development in Drosophila melanogaster. However, the functions of Crinkled have not been fully investigated. Additionally, the genes that participate in the Crinkled pathway are unknown. Phylogenetic analysis indicates that crinkled exhibits a one-to-one orthologous relationship in insects. In Tribolium castaneum, the crinkled gene is 6,498 bp in length and consists of six exons. Crinkled expression peaked during two phases in Tribolium: late embryonic and pupal stages. High levels of crinkled mRNA were detected in the fat body, head, epidermis, ovary, and accessory gland of late adults. Knockdown of crinkled using RNA interference (RNAi) severely affected wing morphogenesis in T. castaneum. We further showed that crinkled silencing reduced forked expression through wingless and shaven-baby, and RNAi of forked phenocopied the effects of crinkled knockdown in T. castaneum. This study investigated the development role of crinkled in postembryonic stages and indicated that forked mediates the functions of crinkled during wing morphogenesis in T. castaneum.

Expression analysis of Bombyx mori bidensovirus structural proteins and assembly of virus-like particles in insect cells.

Bombyx mori bidensovirus (BmBDV) is a new designated species of the new genus Bidensovirus in the new family Bidnaviridae, which contains two single-stranded linear DNAs (VD1 and VD2) and causes the chronic densonucleosis disease of silkworm. Previous researches revealed that VD1-ORF3 encodes the major structural proteins VPs. In this work, through western blot, we found that VPs expressed from 48 h post-inoculation and kept increasing until 120 h post-inoculation in midgut of Bombyx mori. In order to further investigate the translation of vp gene, the ORFs (vp1 and vp2) of the VP started just up-stream of the first two candidate initiation codons were expressed in Sf9 cells by a baculovirus expression system. The expression products were purified by gradient density centrifugation and analyzed by Western blot and electron microscopy. The results showed that the expressions of vp1 yielded three proteins (VP1, VP1', and VP2), which are the same with the viral VPs expression in midgut of Bombyx mori, and vp2 generated two VPs with the molecular weights of about 51 kDa (VP2) and 37 kDa. The observation by electron microscopy indicated that these VPs can auto-assemble into virus-like particles that could not be distinguished from virus particles. These findings will provide materials for studying the structure of BmBDV and be helpful in the studies on BmBDV-based disease in silkworms.

The Bombyx mori nucleopolyhedrovirus Bm111 affects virulence but not virus replication.

The Bm111 of Bombyx mori nucleopolyhedrovirus (BmNPV) encodes a small polypeptide (70 amino acids) of which the function remains unknown. To characterize its function, multiple sequence alignments were performed, and the predicted protein was found to share amazingly high (98 %) sequence identity with the Bombyx mandarina nucleopolyhedrovirus ORF110 (Boma110) but negligible with proteins of other insect viruses, indicating the close relationship between these two NPVs with silkworm larvae. The transcription of Bm111 was detected as early as 3 hpi in BmNPV-infected BmN cells, suggesting it is an early gene. To investigate the role of Bm111 in baculovirus life cycle, a Bm111-knockout virus was constructed by bacmid recombination in Escherichia coli. The results showed that knockout of the Bm111 did not affect the replication of virus DNA, but significantly extended the death time of infected silkworm larvae compared to the wild-type or rescued viruses. We also successfully expressed the recombinant protein Bm111 in E. coli to provide sufficient material for subsequent studies. Taken together, our data indicate that Bm111 only affects the virulence of BmNPV, but not its replication.

Study on anti-BmNPV mechanism of branched-chain amino acid aminotransferases in silkworm.

Bombyx mori nucleopolyhedrovirus (BmNPV) is the most important virus that threatens sericulture industry. At present, there is no effective treatment for BmNPV infection in silkworms, and lncRNA plays an important role in biological immune response and host-virus interaction, but there are relatively few studies in silkworms. In this study, the four midgut tissue samples of the resistance strain NB (NB) and susceptible strain 306 (306) and the NB and 306 continuously infected with BmNPV for 96 h are used for whole transcriptome sequencing to analyze the differences in the genetic background of NB and 306 and the differences after inoculation of BmNPV, and the significantly different mRNA, miRNA and lnRNA between NB and 306 after BmNPV inoculation were screened. By comparing NB and 306, 2651 significantly different mRNAs, 57 significantly different miRNAs and 198 significantly different lncRNAs were screened. By comparing NB and 306 after BmNPV inoculation, 2684 significantly different mRNAs, 39 significantly different miRNAs and 125 significantly different lncRNAs were screened. According to the significantly different mRNA, miRNA and lncRNA screened from NB and 306 and NB and 306 after virus inoculation, the mRNA-miRNA-lncRNA regulatory network was constructed before and after virus inoculation, and the BmBCAT-Bomo_chr7_8305-MSTRG.3236.2 regulatory axis was screened from them, and it was found that BmBCAT was not Bomo_chr7_8305 regulated in the genetic background, after viral infection, MSTRG.3236.2 competes for binding Bomo_chr7_8305 regulates BmBCAT. The whole transcriptome sequencing results were verified by qPCR and the time-series expression analysis was performed to prove the reliability of the regulatory network. The BmBCAT-Bomo_chr7_8305-MSTRG.3236.2 regulatory axis may play a potential role in the interaction between silkworms and BmNPV. These results provide new insights into the interaction mechanism between silkworms and BmNPV.

A novel microfluidic chip integrating with microcolumn array electrodes for rapid and ultrasensitive detection of alpha-fetoprotein.

BACKGROUND: Cancer posed a serious threat to human health, and early diagnosis of cancer biomarker was extremely important for the treatment and control of cancer. Electrochemistry-based assays were low-cost, responsive and easy to operate, but there were some challenges in terms of accuracy, detection limit, efficiency and portability. The combination of microfluidic devices and electrochemical methods was expected to construct a high-performance sensing platform, but long-time antigen-antibody incubation was still required. Therefore, a novel microfluidic chip needs to be developed, which has the advantages of good portability, short incubation time, high accuracy, low detection limit and great application to point-of-care testing. RESULTS: A microfluidic sensor based on microcolumn array electrodes was developed, in which microcolumns could create local mixed flow to reduce the incubation time of target molecules and enhance their interaction with the sensing interface. Besides, three dimensional Mxene fibers-gold nanoparticles (3D MF-Au) was modified on the microcolumn array electrodes to increase active sites and provide more electrolyte shuttle holes. The electrolyte turbulence caused by the microcolumn array electrodes could heighten the contact between the target molecules and sensing interface and accelerate the transfer of redox pairs, thus reducing the incubation time of the target molecules and improving the electrochemical responses in synergy with the 3D MF-Au. Herein, the detection of AFP was chosen as a model, and the microfluidic sensor possessed superior performance for analysis of AFP in the range of 0.1 pg mL-1 - 200 ng mL-1 with a low detection limit (LOD) of 0.0648 pg mL-1. SIGNIFICANCE: This microfluidic chip integrating with microcolumn array electrodes has been successfully implemented to detect AFP in human serum, and the results were consistent with that of electrochemical chemiluminescence method. The microfluidic chip provided a new strategy of portability, shortening incubation time and enhancing electrical signals for antigen detection of real samples, which showed great utilization potentiality in point-of-care testing.

Is there a prognostic difference among stage I lung adenocarcinoma patients with different BRAF-mutation status?

BACKGROUND: The data of the prognostic role of V-Raf murine sarcoma viral oncogene homolog B1 (BRAF) mutations in early-stage lung adenocarcinoma (LUAD) patients is scarce. This study aimed to investigate the proportion, clinicopathological features, and prognostic significance of patients with stage I LUAD carrying BRAF mutations. METHODS: We collected 431 patients with pathological stage I LUAD from cBioPortal for Cancer Genomics and 1604 LUAD patients tested for BRAF V600E and epidermal growth factor receptor (EGFR) mutations from Shanghai Pulmonary Hospital. Survival curves were drawn by the Kaplan-Meier method and compared by log-rank test. Cox proportional hazard models, propensity-score matching (PSM), and overlap weighting (OW) were performed in this study. The primary endpoint was recurrence-free survival (RFS). RESULTS: The proportion of BRAF mutations was estimated at 5.6% in a Caucasian cohort. BRAF V600E mutations were detected in six (1.4%) patients in Caucasian populations and 16 (1.0%) patients in Chinese populations. Two BRAF V600E-mutant patients were detected to have concurrent EGFR mutations, one for 19-del and one for L858R. For pathological stage I LUAD patients, BRAF mutations were not significantly associated with worse RFS than wild-type BRAF patients (HR = 1.111; p = 0.885). After PSM and OW, similar results were presented (HR = 1.352; p = 0.742 and HR = 1.246; p = 0.764, respectively). BRAF V600E mutation status also lacked predictive significance for RFS (HR, 1.844; p = 0.226; HR = 1.144; p = 0.831 and HR = 1.466; p = 0.450, respectively). CONCLUSIONS: In this study, we demonstrated that BRAF status may not be capable of predicting prognosis in stage I LUAD patients. There is a need for more data to validate our findings.

Postoperative chemotherapy significantly improves survival of elderly patients with stage IB-II non-small cell lung cancer: A population-based study.

BACKGROUND: There is scant evidence-based information about survival benefits of postoperative chemotherapy in elderly patients with early-stage non-small cell lung cancer (NSCLC). The purpose of this study is to compare the overall survival (OS) and cancer-specific survival (CSS) rates of surgery alone versus postoperative chemotherapy in patients aged ≥70 years with stage I-II NSCLC. METHODS: Elderly patients aged ≥70 years diagnosed with stage I-II NSCLC were selected from the Surveillance, Epidemiology, and End Results (SEER) database from January 1, 2010 to December 31, 2015. OS and CSS were compared between the two groups utilizing overlap weighting analysis, inverse probability of treatment weight (IPTW), and propensity score matching (PSM). RESULTS: Of the 7193 included patients with stage I-II NSCLC who are more than 70 years old, 681 patients (9.5%) received postoperative chemotherapy and 6512 patients (90.5%) received surgery-alone. Median OS was 77 months in postoperative chemotherapy group versus 79 months in surgery-alone group (p = 0.89). The result of IPTW analysis showed the similar results. The probability of patients choosing chemotherapy increased with the AJCC stage and Grade increasing (p < 0.001) and decreased with the growth of age (p < 0.001). The results of subgroup analysis showed that the survival rate of stage IA patients decreased significantly after postoperative chemotherapy (p < 0.01) while the survival rate of stage IB-II patients increased significantly (p < 0.01). At the same time, we found that patients in the postoperative chemotherapy group tended to have better OS than those in the surgery-alone group with the grade and tumor size increasing. CONCLUSION: The results of this study indicated that postoperative chemotherapy could significantly improve the survival of stage IB-II NSCLC patients aged ≥70 years, and decrease the survival of stage IA patients.

Cytotoxicity of AuCu-Cu2S Nanocomposites: Implications for Biological Evaluation of the Nanocomposite Effect on Bombyx mori Silkworms and Cell Lines.

AuCu-Cu2S nanocomposites are unique materials with exceptional properties that have recently received a lot of interest. However, little is known about their potential toxicity in terrestrial organisms and their subsequent effects on the environment. Therefore, it is essential to develop effective methodologies for evaluating AuCu-Cu2S nanocomposites in biological systems. This study reports the biological evaluation of the AuCu-Cu2S nanocomposite from animal and cell entity levels. The Bombyx mori silkworm was used as a model organism to study the effects of different concentrations of AuCu-Cu2S on silkworm development. Transcriptome analysis was also carried out to examine the genetic modulation exerted by the treatment. Moreover, biocompatibility and cytotoxicity of AuCu-Cu2S were evaluated in human bronchial epithelial cells 16HBE, human lung adenocarcinoma, and the insect Spodoptera frugiperda cell sf9 cell lines. The results showed that although AuCu-Cu2S at ≤400 ppm can prolong the eating habit of silkworms and promote the weight of the cocoon layer, there was an increase in silkworm mortality and a decrease in moth formation at a concentration of ≥800 ppm. The genetic regulation by AuCu-Cu2S treatment showed varying effects in the silkworm, primarily related to functions such as transport and catabolism, metabolism of cofactors and vitamins, xenobiotic biodegradation, amino acid, and carbohydrate. 16HBE, PC-9, and sf9 treated with 300 ppm of AuCu-Cu2S showed viability percentages of 60, 20, and 90%, respectively. Thus, AuCu-Cu2S at low concentrations serves as a safe and biocompatible material for the sf9 cell lines but is lethal to 16HBE and PC-9. This research could aid in understanding the biological effects and biocompatibility of AuCu-Cu2S nanocomposites, particularly in the field of biochemistry; however, the mechanisms involved need further exploration.

A portable microfluidic electrochemical sensing platform for rapid detection of hazardous metal Pb2+ based on thermocapillary convection using 3D Ag-rGO-f-Ni(OH)2/NF as a signal amplifying element.

Heavy metal pollution is causing a great threat to ecological environment and public health, which needs an efficient strategy for monitoring. A portable microfluidic electrochemical sensing system was developed for the determination of heavy metal ions. Herein, the detection of Pb2+ was chosen as a model, and a microfluidic electrochemical sensing chip relying on a smartphone-based electrochemical workstation was proposed for rapid detection Pb2+ with the assistance of thermocapillary convection result from the formed temperature gradient. The 3D Ag-rGO-f-Ni(OH)2/NF composites, prepared by one-step hydrothermal method without any Ni precursor salt, were used to further amplify electrochemical signals under the synergistic effect of thermocapillary convection. The thermocapillary convection could accelerate the preconcentration process and shorten the detection time (save 300 s of preconcentration time). The fabricated system exhibited the exceptional competence for monitoring of Pb2+ range from 0.01 µg/L to 2100 µg/L with a low detection limit (LOD) of 0.00464 µg/L. Furthermore, this portable system has been successfully demonstrated for detecting Pb2+ (0.01 µg/L to 2100 µg/L) in river water (LOD = 0.00498 µg/L), fish (LOD = 0.00566 µg/L) and human serum samples (LOD = 0.00836 µg/L), and the results were consistent with inductively coupled plasma-mass spectrometry (ICP-MS). The proposed novel sensing platform provides a cost-effectiveness, rapidly responding and ease-to-use pathway for analysis of heavy metal ions in real samples and shows great potential in point-of-care testing.

Silkworm glycosaminoglycans bind to Bombyx mori nuclear polyhedrosis virus and facilitate its entry.

Interacting with cell surface attachment factors or receptors is the first step for virus infection. Glycans cover a thick layer on eukaryotic cells and are potential targets of various viruses. Bombyx mori nuclear polyhedrosis viruses (BmNPV) is a baculovirus that causes huge economic loss to the sericulture industry but the mechanism of infection is unclear. Looking for potential host receptors for the virus is an important task. In this study, we investigated the role of glycosaminoglycan (GAG) modifications, including heparan sulfate (HS) and chondroitin sulfate (CS), during BmNPV infection. Enzymatic removal of cell surface HS and CS effectively inhibited BmNPV infection and replication. Exogenous HS and CS can directly bind to BmNPV virion in solution and act as neutralizers for viral infection. Furthermore, the expression of enzymes involved in GAG biosynthesis was upregulated in the BmNPV susceptible silkworm after virus administration, but down-regulated in the resistant strain after virus treatment, suggesting that BmNPV was able to utilize host cell machinery to promote the biosynthesis of GAGs. This study demonstrated HS and CS as important attachment factors that facilitate the viral entry process, and targeting HS and CS can be an effective means of inhibiting BmNPV infection.