Exploring the key deteriorative microorganisms on ancient ivories unearthed from the Sanxingdui Ruins site during temporary cold storage.

Introduction: The ancient ivories unearthed from the Sanxingdui Ruins site are valuable cultural relics, however, the microbial biodeterioration on ivories during temporary cold storage poses a great threat to their later long-term preservation. Methods: Here, the combination of high-throughput sequencing and biochemical assays was applied for the in-depth investigation of the key deteriorative microorganisms colonizing on the ivories and the tracing of their origin, as well as the assessment of the ethanol disinfection impact on the microbial communities on ivories. Results: It was observed that the surfaces of ivories were scattered by the fungal patches of white, dark grey, and hedge green colors during cold storage. The high-throughput sequencing results showed that the genera Mortierella (38.51%), Ilyonectria (14.43%), Penicillium (1.15%), and Aspergillus (1.09%) were the dominant fungi, while Pseudomonas (22.63%), Sphingopyxis (3.06%), and Perlucidibaca (2.92%) were the dominant bacteria on ivories. The isolated Aspergillus A-2 resulted in the highest amount of calcium releasing from the degradation of hydroxyapatite (HAP), the main component of ivory, by the organic acids produced, including oxalic acid and citric acid. The fast expectation-maximization for microbial source tracking (FEAST) analysis revealed that the majority of the fungi (57.45%) and bacteria (71.84%) colonizing on the ivories were derived from the soils surrounding ivories in the sacrifice pits, indicating soils as the primary source for the spoilage microbes growing on ivories. The dominant strains could degrade cellulose, the key components of wet cotton towels commonly applied on ivories for moisture maintenance, aiding the spoilage microbes colonizing on ivories. Notably, the ivory disinfection with 75% ethanol during the cleansing significantly decreased the relative abundance of the dominant genera of Ilyonectria, Aspergillus, and Pseudomonas, with Mortierella becoming the dominant one on ivories. Discussion: Together, the fungi, particularly Aspergillus and Penicillium, played a significant role in the microbial biodeterioration of unearthed ancient ivories by producing the organic acids. These results may improve the control of the microbial biodeterioration and develop more efficient strategies for the long-time conservation of unearthed ancient ivories and other cultural relics.

Proteomic investigation and understanding on IgY purification and product development.

An increasing demand for the development of immunoglobin Y (IgY) illustrates the necessity of the component analysis in the process of conduction and quality control. This study investigated the proteomic changes in crude IgY extracts and purified IgY products obtained by sequential polyethylene glycol precipitation (PEG) of egg yolks followed by human mycoplasma protein-based affinity chromatography compared with intact egg yolks. After confirming the extraction efficiency and purity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, liquid chromatography tandem-mass spectrometry (LC-MS/MS) was performed with samples including fresh yolk, IgY extracted product and purified product. A total of 348 proteins were identified, with 36 proteins deleted and 209 newly detected proteins in the purified product compared to the intact egg yolk. The significantly decreased proteins mainly included phosvitin, albumin, and apolipoprotein B whereas the significantly increased proteins were mainly IgY-related proteins. GO analysis showed that the purified IgY product had ATPase activity and purine ribonucleoside triphosphate binding activity, and was mainly involved in purine and nucleic acid metabolism. This study will inevitably fasten the commercial application of IgY antibodies and is of greater significance for promotion, development and approval for new antibody derived drug products.

Trends in industrialization and commercialization of IgY technology.

IgY technology refers to the strategic production process involved in generating avian immunoglobulin (IgY) against target antigens in a much more cost-effective manner with broad applications in the fields of diagnostics, prophylaxis, and therapeutics for both human and veterinary medicine. Over the past decade, promising progress in this research area has been evident from the steep increase in the number of registered manufacturing companies involved in the production of IgY products, the number of patents, and the notable number of clinical trials underway. Hence, it is crucial to conduct a prospective analysis of the commercialization and marketing potential of IgY-based commercial products for large-scale applications. This review revealed that the number of IgY patent applications increased steeply after 2010, with the highest of 77 patents filed in 2021. In addition, 73 industries are reportedly involved in marketing IgY products, out of which 27 were promoting biotherapeutics for human and veterinary medicine and 46 were in the diagnostic field. IgY antibodies are being used as primary and secondary antibodies, with approximately 3729 and 846 products, respectively. Biotherapeutic product consumption has notably increased as a food supplement and as a topical application in human and veterinary medicine, which are under different clinical phases of development to reach the market with around 80 and 56 products, respectively. In contrast, the number of IgY products as parenteral administrations and licensed drugs is not well developed given the lack of technical standards established for IgY registration and industrialization, as well as the restriction of the nature of polyclonal antibodies. However, recent ongoing research on functional IgY fragments indicates a promising area for IgY applications in the near future. Therefore, retrospective analysis with speculations is mandatory for IgY technology maturation toward industrialization and commercialization.

Label-Free LC-MS/MS Analysis Reveals Different Proteomic Profiles between Egg Yolks of Silky Fowl and Ordinary Chickens.

The proteomic profiles of Silky fowl egg yolk (SFEY) and Leghorn egg yolk (LEY) were analyzed by bottom-up label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). From a total of 186 identified proteins, 26 proteins were found significantly differentially abundant between two yolks, of which, 19 were up-regulated and 7 were down-regulated in SFEY, particularly, vitelline membrane outer layer protein 1, transthyretin and ovoinhibitor were up-regulated by 26, 25, and 16 times, respectively. In addition, there were 57 and 6 unique proteins in SFEY and LEY, respectively. Gene Ontology (GO) revealed SFEY contained relatively more abundant protease inhibitors and coagulation-related proteins. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed differentially abundant proteins in SFEY may be actively involved in the regulation of the neuroactive ligand-receptor interaction pathway. This study provides a theoretical basis for the understanding of proteomic and biological differences between these two yolks and can guide for further exploration of nutritional and biomedical use of Silky fowl egg.

Specific anti-SARS-CoV-2 S1 IgY-scFv is a promising tool for recognition of the virus.

As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread globally, a series of vaccines, antibodies and drugs have been developed to combat coronavirus disease 2019 (COVID-19). High specific antibodies are powerful tool for the development of immunoassay and providing passive immunotherapy against SARS-CoV-2 and expected with large scale production. SARS-CoV-2 S1 protein was expressed in E. coli BL21 and purified by immobilized metal affinity chromatography, as antigen used to immunize hens, the specific IgY antibodies were extracted form egg yolk by PEG-6000 precipitation, and the titer of anti-S1 IgY antibody reached 1:10,000. IgY single chain variable fragment antibody (IgY-scFv) was generated by using phage display technology and the IgY-scFv showed high binding sensitivity and capacity to S1 protein of SARS-CoV-2, and the minimum detectable antigen S1 protein concentration was 6 ng/µL. The docking study showed that the multiple epitopes on the IgY-scFv interacted with multiple residues on SARS-CoV-2 S1 RBD to form hydrogen bonds. This preliminary study suggests that IgY and IgY-scFv are suitable candidates for the development of immunoassay and passive immunotherapy for COVID-19 to humans and animals.

Scientometric analysis and perspective of IgY technology study.

Egg yolk immunoglobulin (IgY) is a class of antibody that is produced in birds against pathogens. Therefore, hyperimmunization of birds can produce a specific antibody in the egg against target antigen for a wide range of applications in diagnostic, prophylactic or treatment in human and veterinary medicine which is known today as IgY technology. Until now, the number of articles, patents and clinical studies on IgY technology has increased significantly. Hence, there is a fact that scientometric studies are needed to gain a deeper understanding of the research for the commercialization of IgY technology. Until now, no scientometric research has been directed toward IgY technology. In view of this, we conducted scientometric analysis in the WoS database. A total of 1,029 IgY-related papers were obtained including 981 journal articles and 48 reviews. The visualization of this literature showed an increasing trend in the number of IgY-related publications over the 4 decades, especially after 2008 to 2021. China, the United States, Canada, Japan, and Germany had the largest number of publications, with 220, 148, 91, 76, and 72, respectively. Among all the research institutions, Dalian University of Technology, Alberta University and Northwest Agriculture and Forestry University published the most of the articles, respectively. Among authors, Dr. Xiaoying Zhang had the highest number of publications with 21. The top most cited publications were from Dr. da Silva with 38 citations. Keywords co-occurrence network analysis showed that the correlation between different keywords is large, especially IgY, antibodies and immunoglobulin which is consistent with the rapid increase in the number of publications. Finally, through this data analysis, we hope that our result could help IgY technology to more maturity toward industrialization and commercialization.

Emodin inhibits the expression of receptor and calcitonin-gene-related peptide release in trigeminal ganglia of trigeminal neuralgia rats.

Trigeminal neuralgia (TN) is one of the most intense forms of facial pain. It has been reported that the P2X3 receptor plays a crucial role in facilitating pain transmission, and the calcitonin-gene-related peptide (CGRP) from trigeminal ganglia (TGs) might perform differing function in nociceptive afferent input transmission. The present study investigated whether emodin can affect TN pain transmission by suppressing the expression of P2X3 receptors and CGRP in TGs. Chronic constriction injury of the infraorbital branch of the trigeminal nerve (CCI-ION) was used as TN model. The TN rats were randomly divided into the following 4 groups: (1) a sham group (Sham), (2) a sham rats treated with emodin group (TN + E), (3) a TN rats treated with 0.5% sodium carboxymethyl cellulose (CMC) as vehicle group (TN) and (4) a TN rats treated with emodin group (TN + E). The mechanical hyperalgesia threshold of TN rats was tested by Electric Von Frey filaments. The change of the expression of P2X3 receptors and CGRP in rat's TG was detected with RT-PCR, immunohistochemical staining, and Western blotting. The phosphorylation of p38 and ERK1/2 pathway of TG was detected by Western blotting. After CCI-ION injury, the threshold of mechanical hyperalgesia for the territory of ligated infraorbital nerve in TN group decreased significantly compared with that in sham group. On day 14 after operation of CCI-ION, there was also an evident increase in the expression of P2X3 receptors and CGRP in the TG of TN group. However after treatment with emodin, the response of mechanical hyperalgesia of TN rats was clearly increased while the enhanced expression of P2X3 receptor and CGRP in TN rats was significantly decreased. The phosphorylation of p38 and ERK1/2 in TN group was stronger than that in Sham group. But these phosphorylation changes in the TN rats were much weaker after treatment with emodin. In conclusion, P2X3 receptor may cooperate with CGRP in the pain transmission of TN, and emodin can inhibit the expression and activation of P2X3 receptor and CGRP in TG to relieve TN.

Simultaneous determination of phenolic compounds in sesame oil using LC-MS/MS combined with magnetic carboxylated multi-walled carbon nanotubes.

A novel magnetic carboxylated multi-walled carbon nanotubes (c-MWCNT-MNPs) was proposed for magnetic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry to determine phenolic compounds in sesame oil. In this study, c-MWCNT-MNPs were acquired by simply dispersing Fe3O4 magnetic nanoparticles into carboxylated multi-walled carbon nanotubes. The major parameters affecting extraction efficiency were optimized, including the type and volume of desorption solvents, extraction and desorption time, washing solution, and sorbent amount. The limit of quantifications and limit of detections were from 0.03µg/kg to 43.00µg/kg and from 0.01µg/kg to 13.60µg/kg, respectively. The recoveries of phenolic compounds in vegetable oils were in the range of 83.8-125.9% with inter-day and intra-day precisions of less than 13.2%. It was confirmed that this method was simple, rapid and reliable with an excellent potential for routine analysis of phenolic compounds in oil samples.

[Application of magnetic resonance diffusion weighted imaging in early ankylosing spondylitis].

OBJECTIVE: To evaluate the magnetic resonance diffusion-weighted imaging (DWI) in the detection of early ankylosing spondylitis and explore the manifestations of ankylosing spondylitis on whole body DWI (WB-DWI). METHODS: A total of 16 patients with early ankylosing spondylitis (AS) and 18 patients with low back pain (LBP) were recruited. Subchondral bone marrow apparent diffusion coefficient (ADC) in bilateral ilium and sacrum along sacroiliac joints were compared. An independent sample t-test (SPSS 16.0, SPSS, Chicago, III) was utilized to analyze the ADC value differences between groups. P < 0.05 denoted statistical significance. The mean ADC values of focal lesions in AS patients were also measured. Whole body diffusion weighted imaging was performed in additional 8 clinical confirmed AS patients and analyzed with the techniques of maximum intensity projection (MIP) and multiplanar reconstruction (MPR) in comparison with conventional MRI images to investigate the detectability of AS lesions with whole body DWI. RESULTS: Mean ADC values in 16 AS patients were (0.51 ± 0.13)×10(-3)mm(2)/s in ilium and (0.49 ± 0.17)×10(-3)mm(2)/s in sacrum. Mean ADC values in 18 LBP patients were (0.32 ± 0.06)×10(-3)mm(2)/s in ilium and (0.31 ± 0.08)×10(-3)mm(2)/s in sacrum. The ADC value in AS patients were statistically significantly greater than those in ilium and sacrum of LBP patients. Whole body DWI detected abnormalities in 8 AS patients within bilateral sacroiliac joints and other sites corresponding to the clinical symptoms of patients. The mean ADC values of focal lesions of this patient cohort were (1.31 ± 0.38)×10(-3)mm(2)/s in sacrum and (1.18 ± 0.27)×10(-3)mm(2)/s in ilium. CONCLUSION: Subchondral marrow ADC values along sacroiliac joints allow the differentiation of patients with early AS from LBP patients. In conjunctions with such post-processing techniques as MIP and MPR, WB-DWI allows a comprehensive assessment of AS patients to guide treatment, evaluate prognosis and follow therapeutic responses.

A new alternative for bony chest wall reconstruction using biomaterial artificial rib and pleura: animal experiment and clinical application.

OBJECTIVE: To evaluate a new method for chest wall reconstruction using porcine-derived artificial rib and pleura in an animal experiment. Further, the clinical application was performed in five patients with large defects in the chest wall as a preliminary observation. METHODS: In animal experiments, a full-thickness chest wall defect of 7 cm × 8 cm was created in 12 adult mongrel dogs. Six dogs underwent reconstruction with porcine-derived artificial ribs and pleura (test group), and six with methylmethacrylate and double polyester mesh in the form of traditional Marlex sandwich technique (control group). At follow-up of each for 3, 6, and 12 months postoperatively, a general performance assessment and thoracic radiography were performed. Gross and histopathological examinations were carried out following humane euthanasia at the time of last follow-up. In clinical application, five patients with wide tumor resection in the chest wall underwent reconstruction with porcine-derived artificial ribs and pleura as well. RESULTS: In animal experiment, no perioperative death or hyperpyrexia occurred and no difference in either infection or dyspnea was noted between the two groups. Postoperative radiography revealed good thoracic integrity with no evidence of collapse, deformation, or abnormal movement in the test group. In the control group, similar results were observed, except that two dogs had abnormal movement in the chest wall associated with respiration. Severe adhesions between the 'sandwich' complex and the host tissues were identified in the control group, but by contrast, only mild adhesions were noted in the test group. The non-degradable polyester mesh induced fibrous proliferation and rejection, whereas the artificial pleura was absorbed with mild fibrous hyperplasia after 12 months. In clinical application, no thoracic deformity, chronic pain, or respiratory discomfort were observed at 1 or 12 postoperative months. CONCLUSIONS: Porcine-derived ribs and pleura can be employed safely to create an artificial chest wall to repair bony chest defects. The clinical results corresponded well with those of animal experiments, and thus confirmed the safety and feasibility of this new alternative of chest wall reconstruction. However, a long-term study in a large number is needed due to the small number of animals in this study.

Effects of rat serum containing Chinese herbal medicine Sangen Decoction on osteoclastogenesis and bone resorption of osteoclasts induced by polymethylmethacrylate particles / 中西医结合学报

To investigate the effects of Sangen Decoction, a compound Chinese herbal medicine, on osteoclastogenesis and bone resorption function of osteoclasts induced by polymethylmethacrylate particles in vitro.

Effect of ZGDHu-1 on proliferation and apoptosis of A549 cells in vitro and antitumor activity in vivo / 药学学报

This study is to explore the mechanism and effect of N, N'-di-(m-methylphenyl)-3, 6-dimethyl-1, 4-dihydro-1, 2, 4, 5-tetrazine-1, 4-dicarboamide (ZGDHu-1) on proliferation and apoptosis of A549 cells in vitro and on A549 xenograft tumor in nude mice. With different concentrations of ZGDHu-1 at different times were used to treat A549 cells in vitro. The proliferation was determined by living cell count, SRB assay and Brdu-ELISA. Cell apoptosis was determined by cell morphology, DNA agarose gel electrophoresis, DNA content, Annexin V/PI and Hoechst 33258 labeling method. The nude mice model of A549 xenograft tumor was established by subcutaneous inoculation. The suppression activity of ZGDHu-1 by intraperitoneal injection on xenograft mice model was detected. The expressions of bcl-2, bax and p53 gene and protein were analyzed by RT-PCR and flow cytometry. ZGDHu-1 can inhibit A549 cell proliferation viability within a certain range of treating time and does, and a majority of A549 cells were arrested in G2-M phase. The A549 cells apoptosis was confirmed by typical cell morphology, DNA fragment, Sub G1 phase, Hoechst 33258 and Annexin V/PI labeling method with a time and dose related manner. When the xenograft tumor mice model were treated with 10, 20 and 40 mg x kg(-1) ZGDHu-1 for 14 days, the tumor growth inhibition rate were 43.7%, 56.9% and 60.0%, respectively. The expression of bax, bax/bcl-2 and p53 gene and protein increased significantly and bcl-2 decreased slightly by the treatment of ZGDHu-1. ZGDHu-1 can significantly suppress the growth of A549 xenograft tumor in vivo and inhibited proliferation by inducing tumor cell apoptosis in vitro. The mechanism may associate with its up-regulation of bax and p53 during the apoptosis process.