Effect of Symptom Levels of Children's Food Allergy on Maternal Depression: A Cross-sectional and Cohort Study.

BACKGROUND: Maternal depression may have negative impacts on children's behavior and mental health. Childhood food allergy is a common health issue, yet its relationship with maternal depression remains incompletely understood. This study aimed to analyze the association between children's food allergy symptoms and maternal depression through cross-sectional and cohort studies. METHODS: This study selected a total of 580 children with food allergy and their mothers who met the inclusion criteria in Ganzhou Women and Children's Health Care Hospital from April 2015 to April 2022, evaluated the symptom levels of children's food allergy according to the guidelines, assessed the depressive symptoms of mothers using self-rating depression scale (SDS), and analyzed the relationship between the symptom severity of children's food allergy and the risk of maternal depression; at the same time, one-year follow-up of mothers without depression was carried out to measure the incidence of depression to further explore this relationship. RESULTS: The 580 children with food allergies in the cross-sectional study consisted of 365 (62.93%) males and 215 (37.07%) females, aged (8.98 ± 2.30) years, with 298 (51.37%) experiencing Level-Ⅰ, and 282 (48.63%) experiencing Level-Ⅱ. A total of 56 (9.66%) mothers suffered from depression, aged (42.74 ± 5.42) years. Adjusting for confounders including mother's age, education level, marital status, family income, comorbidities, history of allergies, family history of food allergies, history of psychiatric disorders, current smoking status, current alcohol consumption, current regular exercise status, childhood food allergens and food allergy categorization, the mothers of children with child food allergy symptom Level-Ⅱ were found to have a higher risk of depression compared with mothers with child food allergy symptom Level-Ⅰ, odds ratio (OR) = 2.025 (95% confidence interval (CI): 1.319-3.128, p = 0.001). In the one-year cohort study, 38 (7.25%) mothers had new-onset depressive symptoms. Mothers of children with a child food allergy symptom Level-Ⅱ had an OR = 2.165 (95% CI: 1.612-2.902, p < 0.001) for depressive symptoms compared to mothers with a child food allergy symptom Level-Ⅰ. CONCLUSION: Among children with food allergy symptom scores of Level-Ⅰ and Level-Ⅱ, higher levels were associated with a higher prevalence of depression in their mothers.

Solvent-Free Enzymatic Synthesis of Ethyl 3-Oxobutyrate Derivative: Characterization and Optimization of Reaction Condition.

It was first found that porcine pancreatic lipase (PPL) could catalyze the Knoevenagel condensation of aromatic aldehydes and ethyl acetoacetate under solvent-free conditions in this paper. Under solvent-free conditions, the highest yield of PPL catalytic reaction was 99.38%, and the Z/E selectivity of the product was 3.93. In addition, the reaction conditions were optimized, and the factors affecting the product structure were studied.

Nano Carbon Black-Based High Performance Wearable Pressure Sensors.

The reasonable design pattern of flexible pressure sensors with excellent performance and prominent features including high sensitivity and a relatively wide workable linear range has attracted significant attention owing to their potential application in the advanced wearable electronics and artificial intelligence fields. Herein, nano carbon black from kerosene soot, an atmospheric pollutant generated during the insufficient burning of hydrocarbon fuels, was utilized as the conductive material with a bottom interdigitated textile electrode screen printed using silver paste to construct a piezoresistive pressure sensor with prominent performance. Owing to the distinct loose porous structure, the lumpy surface roughness of the fabric electrodes, and the softness of polydimethylsiloxane, the piezoresistive pressure sensor exhibited superior detection performance, including high sensitivity (31.63 kPa-1 within the range of 0-2 kPa), a relatively large feasible range (0-15 kPa), a low detection limit (2.26 pa), and a rapid response time (15 ms). Thus, these sensors act as outstanding candidates for detecting the human physiological signal and large-scale limb movement, showing their broad range of application prospects in the advanced wearable electronics field.

Developing and validating utility parameters to establish patient-reported outcome-based perioperative symptom management in patients with lung cancer: a multicentre, prospective, observational cohort study protocol.

INTRODUCTION: Patient-reported outcome-based symptom monitoring and alerting have been attractive for patient care after a tumour-removal surgery. However, the implementation parameters of this patient-centred symptom management system in perioperative patients with lung cancer are still lacking. We aim to develop a perioperative symptom scale (PSS) for monitoring, to determine the optimal time points for symptom assessment and to define the alert thresholds for medical intervention. METHODS AND ANALYSIS: This study will prospectively recruit 300 patients undergoing lung cancer surgery in six hospitals. The MD Anderson Symptom Inventory-Lung Cancer Module (MDASI-LC) is used to collect longitudinal symptom data preoperatively, daily postoperatively during in-hospital stay and weekly after discharge until 4 weeks or the start of postoperative oncological therapy. Symptoms that change significantly over time will be generated as the PSS. We will determine the optimal time points for follow-up using the generalised linear mixed-effects models. The MDASI-LC interference-measured functional status will be used as the anchor for the alert thresholds. ETHICS AND DISSEMINATION: Ethics Committee of Sichuan Cancer Hospital approved this study on 16 October 2017 (No. SCCHEC-02-2017-042). The manuscript is based on the latest protocol of Version 3.0, 15 September 2019. The results of this study will be presented at medical conferences and published in peer-reviewed journals. TRIALS REGISTRATION NUMBER: NCT03341377.

Using patient-reported outcomes to manage postoperative symptoms in patients with lung cancer: protocol for a multicentre, randomised controlled trial.

INTRODUCTION: Surgery is one of the primary treatments for lung cancer. The postoperative symptom burden experienced by patients with lung cancer is substantial, seriously delaying their recovery from surgery and impairing their quality of life. Patient-reported outcome (PRO)-based symptom management is increasingly regarded as an optimal model for patient-centred care. Currently, clinical trial-based evidence involving early-phase (immediately after surgery for up to 1 month) symptom management of lung cancer is lacking. We propose a randomised trial to evaluate the effect of a PRO-based symptom-monitoring programme with overthreshold alerts and responses for postoperative recovery in patients with lung cancer. METHODS AND ANALYSIS: The study will recruit 160 patients with lung cancer from six hospitals. The patients will be randomly allocated to the intervention group or control group in a ratio of 1:1. Patients in the intervention group will receive PRO-based symptom management from the specialists when their reported target symptom (pain, coughing, fatigue, disturbed sleep and shortness of breath) scores reach the preset threshold (score ≥4). Patients in the control group will not generate alerts and will follow the standard procedures for symptom management. All patients will receive symptom assessments via the MD Anderson Symptom Inventory-lung cancer module on the day before surgery, daily after surgery and twice a week after discharge until 4 weeks or the start of postoperative oncological treatment. The primary outcome-mean symptom threshold events-will be compared between the intervention and control group via independent sample Student's t-test. ETHICS AND DISSEMINATION: The study was approved by the Ethics Committee of Sichuan Cancer Hospital on 22 November 2018 (No. SCCHEC-02-2018-045). This manuscript is based on V.2.0, 9 May 2019 of the protocol. The study results will be disseminated in publications in peer-reviewed journals and presentations at academic conferences. TRIALS REGISTRATION NUMBER: ChiCTR1900020846.

Investigation of Energy Transfer in Star-Shaped White Polymer Light-Emitting Devices via the Time-Resolved Photoluminescence.

A series of white polymer light-emitting devices (WPLEDs) were fabricated by utilizing star-shaped white-emission copolymers containing tri[1-phenylisoquinolinato-C2,N]iridium (Ir(piq)3), fluorenone (FO) and poly(9,9-dioctylfluorene) (PFO) as red-, green- and blue-emitting (RGB) components, respectively. In these WPLEDs, a maximum current efficiency of 6.4 cd·A-1 at 20 mA·cm-2 and Commission Internationale d'Eclairage (CIE) coordinates of (0.33, 0.32) were achieved, and the current efficiency was still kept to 4.2 cd·A-1 at the current density of 200 mA·cm-2. To investigate energy transfer processes among the three different chromophores of the star-shaped copolymers in these WPLEDs, the time-resolved photoluminescence (PL) spectra were recorded. By comparing the fluorescence decay lifetimes of PFO chromophores in the four star-like white-emitting copolymers, the efficient energy transfer from PFO units to Ir(piq)3 and FO chromophores was confirmed. From time-resolved PL and the analysis of energy transfer process, the results as follows were proved. Owing to the star-like molecular structure and steric hindrance effect, intermolecular interactions and concentrations quenching in the electroluminescence (EL) process could also be sufficiently suppressed. The efficient energy transfer also reduced intermolecular interactions' contribution to the enhanced device performances compared to the linear single-polymer white-light systems. Moreover, saturated stable white emission results from the joint of energy transfer and trap-assisted recombination. This improved performance is expected to provide the star-like white-emitting copolymers with promising applications for WPLEDs.

Orf Virus Encoded Protein ORFV119 Induces Cell Apoptosis Through the Extrinsic and Intrinsic Pathways.

Apoptosis, a significant form of cell death, has a leading role in the host cell defense against virus infection. Viruses have evolved a series of strategies that block apoptosis during the early stage of viral infection to enhance viral replication, and induce apoptosis in the late stages to facilitate viral particle release from the cells. Here we show that orf virus (ORFV), the causative agent of orf, encodes an apoptosis-inducing protein ORFV119. ORFV119 targets the mitochondria in host cells, inhibits cell proliferation, and induces cell apoptosis. Protein array data indicated that ORFV119 could induce apoptosis via up-regulation of Smac, Bak, and Bax and down-regulation of anti-apoptotic proteins Bcl-2 and cIAP-2. Activation of caspase-9 and caspase-3, and consequent PARP cleavage, ultimately lead to apoptosis. ORFV119 could also directly activate caspase-8 and induce Bid, involved in the extrinsic pathway, to achieve cell death. Furthermore, sequence analysis and experiments with mutants of ORFV119 introduced revealed that ORFV119 contains a key N-terminal domain that is necessary and sufficient to direct the protein to the mitochondria. Together, we report, for the first time, the identification of the novel apoptosis-inducing protein ORFV119 encoded by a parapoxvirus. This provides an important reference for the study of pathogenesis, identification of immunomodulation mechanisms of ORFV, and may lead to new strategies for orf disease control.

Baicalein Accelerates Tendon-Bone Healing via Activation of Wnt/ß-Catenin Signaling Pathway in Rats.

BACKGROUND: Tendon-bone healing is a reconstructive procedure which requires a tendon graft healing to a bone tunnel or to the surface of bone after the junction injury between tendon, ligament, and bone. The surgical reattachment of tendon to bone often fails due to regeneration failure of the specialized tendon-bone junction. MATERIALS AND METHODS: An extra-articular tendon-bone healing rat model was established to discuss the effect of the baicalein 10 mg/(kg·d) in accelerating tendon-bone healing progress. Also, tendon-derived stem cells (TDSCs) were treated with various concentrations of baicalein or dickkopf-1 (DKK-1) to stimulate differentiation for 14 days. RESULTS: In vivo, tendon-bone healing strength of experiment group was obviously stronger than the control group in 3 weeks as well as in 6 weeks. And there were more mature fibroblasts, more Sharpey fibers, and larger new bone formation area treated intragastrically with baicalein compared with rats that were treated with vehicle for 3 weeks and 6 weeks. In vitro, after induction for 14 days, the expressions of osteoblast differentiation markers, that is, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), osterix (OSX), and collagen I, were upregulated and Wnt/ß-catenin signaling pathway was enhanced in TDSCs. The effect of DKK-1 significantly reduced the effect of baicalein on the osteogenic differentiation. CONCLUSION: These data suggest that baicalein may stimulate TDSCs osteogenic differentiation via activation of Wnt/ß-catenin signaling pathway to accelerate tendon-bone healing.

[Osteopractic total flavone promoting rat extra-articular tendon-bone healing through mTOR pathway].

OBJECTIVE: To explore function and related molecular mechanism of osteopractic total flavone (OTF) on tendon healing in rats. METHODS: Ten male rats aged for 8 weeks were collected and weighted from 180 to 220 g. Tendon stem cells were cultivated, the third tendon stem cells were used for experiment. OTP treated with 0, 0.1, 1, 10 ng/ml were added into tendon stem cells, and expression change of ALP, Runx2, OCN, VEGF, P-S6, P-4E/BP1 were detected after 14 days. Forty male rats aged for 8 weeks (weighted 180 to 220 g) were established extra-articular tendon-bone transplanting healing model, and divided into experimental group and control group. Experimental group were treated with OTF(100 mg·kg⁻¹·d⁻¹), while control group was treated by normal saline with the same volume. Tendon-bone healing degree were detected by biomechanical testing at 3 and 6 weeks after surgery, histological detection were applied to detect tendon-bone healing and number of new vessles. RESULTS: After treated by OTP, ALP staining and active index detection showed there were statistical differences among 0, 0.1, 1, 10 ng/ml group. After 14 days' cultivation, western blotting results showed mTOR downstream marker protein P-S6 protein expression were gradually increased with increase of density of OTP, expression of P-4E/BP1 was reduced, while expression of Runx2, OCN, VEGF were increased. Biological detection results showed that there was no significant difference in mechanical strength between experimental group(0.78±0.05) N/mm and control group (0.51±0.02) N/mm at 3 weeks after surgery, while mechanical strength in experimental group (1.36±0.09) N/mm was higher than control group (1.01±0.08) N/mm at 6 weeks after surgery. Histological results showed maturity of tendon-bone surface cell were higher at 3 and 6 weeks in experimental group, sharpey fiber growth more density, calcification extent of mesenchyme was high, and new bone, vessels were increased. CONCLUSIONS: OTF could promote osteogenic differentiation of tendon stem cells through mTOR signaling in vitro, and stimulate tendon-bone healing in bone tunnel and enhance connection quality between tendon and bone.

Solution-Processed Warm White Organic Light-Emitting Diodes Based on a Blue Thermally Activated Delayed Fluorescence Dendrimer.

Thermally activated delayed fluorescence (TADF) emitters have received much attention for the fabrication of white organic light-emitting diodes (WOLEDs); however, challenges remain owing to severe efficiency roll-off, poor color stability, and high cost. In this contribution, solution-processed hybrid WOLEDs were obtained by employing a blue TADF dendrimer, bis{4-[3,6-bis(3,6-di-tert-butylcarbazol-N-yl)carbazol-N-yl]phenyl}sulfone (BPS), combined with an orange iridium complex, bis(2-phenylbenzothiazolato)(acetylacetonate)iridium(III), Ir(bt)2 (acac), as a co-dopant. The devices achieved a maximum external quantum efficiency of 6.59 % and a maximum current efficiency of 17.34 cd A-1 . The results suggest that the TADF dendrimer serving as an assistant dopant were helpful in reducing the triplet populations by up-converting the triplet excitons to the singlet state and immediately transferring the singlet excitons to Ir(bt)2 (acac) (bt=2-phenylbenzothiazolato, acac=acetylacetonato) by virtue of the long-range Förster resonance energy transfer, thus significantly decreasing the triplet-triplet annihilation (TTA). Moreover, the emitters can act as shallow trapping centers to decrease charge and exciton aggregation. The resulting OLEDs exhibit stable electroluminescent spectra and low efficiency roll-off at relatively high current density. The CIE coordinates Δ(x, y) vary only (0.02, 0.02) in the luminance range of 100 to 10 000 cd m-2 .

The whole genomic analysis of orf virus strain HN3/12 isolated from Henan province, central China.

BACKGROUND: The Orf virus (ORFV) is the causative agent of orf, a globally-occurring, acute, pustular, contagious disease affecting sheep, goats and humans with a worldwide distribution. Currently, the genomic analysis of four ORFV strains from the Fujian province in southern China and a NA1/11 strain isolated from the Jilin province in northeast China have been reported. However, little is known about the genomic information of ORFV strains from central China. RESULTS: From a recent outbreak in a sheep herd in the Henan province of central China, a novel ORFV strain (HN3/12) was isolated and cultured in ovine fetal turbinate (OFTu) cells. The strain was identified as HN3/12 and verified by PCR based on the DNA sequences of 011 and 059 genes. The whole genomic sequence of this isolate was determined by Next Generation Sequencing technology. To determine the genetic characteristics of the HN3/12 strain, phylogenetic analysis of the 011 and 059 genes and amino acid sequence alignment of the HN3/12 strain were performed and compared with reference parapoxvirus strains. CONCLUSIONS: The HN3/12 genome is 136,643 bp in length, contains 63.67% G + C and encodes 132 putative genes. Phylogenetic analysis of the 011 and 059 nucleotide sequences showed that this viral strain was similar to the NA1/11 isolate. The homology analysis indicates that HN3/12 has 93% to 98% identity with published ORFV strains at amino acid level. When open reading frames (ORFs) were aligned among the HN3/12 and four Fujian ORFV strains, most of them have identities greater than 90% and only a few less than 60%. The availability of the whole genomic sequence of HN3/12 aids in our understanding of, and provides new insights into, the genetic diversity of ORFV.

Extracellular translationally controlled tumor protein promotes colorectal cancer invasion and metastasis through Cdc42/JNK/ MMP9 signaling.

The translationally controlled tumor protein (TCTP) can be secreted independently of the endoplasmic reticulum/Golgi pathway and has extrinsic activities when it is characterized as the histamine releasing factor (HRF). Despite its important role in allergies and inflammation, little is known about how extracellular TCTP affects cancer progression. In this study, we found that TCTP was overexpressed in the interstitial tissue of colorectal cancer (CRC) and its expression correlated with poor survival, high pathological grades and metastatic TNM stage in CRC patients. TCTP expression was greater in metastatic liver tissue than in primary tumors and was increased in highly invasive CRC cells. We demonstrated that the expression of TCTP was regulated by HIF-1α and its release was increased in response to low serum and hypoxic stress. Recombinant human TCTP (rhTCTP) promoted the migration and invasiveness of CRC cells in vitro and contributed to distant liver metastasis in vivo. Furthermore, rhTCTP activated Cdc42, phosphorylated JNK (p-JNK), increasing the translocation of p-JNK from the cytoplasm to the nucleus, as well as the secretion of MMP9. In addition, the expression of TCTP positively correlated with that of Cdc42 and p-JNK in clinical CRC samples. The silencing of Cdc42, JNK and MMP9 significantly inhibited the Matrigel invasion of rhTCTP-enhanced CRC cells. Collectively, these results identify a new role for extracellular TCTP as a promoter of CRC progression and liver metastases via Cdc42/JNK/MMP9 activation.

[Preparation,Identification,and Application of Monoclonal Antibody against Orf Virus 118 Protein].

This study was designed to prepare a monoclonal antibody against Orf virus 118 protein and explore the biological properties of ORFV118 using this antibody. We constructed a recombinant plasmid pET33b-ORFV118 that contained a full-length ORFV118 gene. The plasmid was transformed into E.coli BL21,and the expression of ORFV118 was induced by isopropyl-ß-d-thiogalactoside (IPTG).Prokaryotic ORFV118 was purified via Ni-NTA affinity chromatography and was subsequently used as an antigen to immunize mice. An anti-ORFV118 antibody was prepared using hybridoma technology. The titer and specificity of this antibody were tested by an indirect ELISA and Western blot/immunohistochemistry, respectively. We successfully obtained three antibody-secreting hybridomas,1A2,3B5,and 5D10.The titers of the three hybridomas were 1:10000,1:6400,and 1:8000.The monoclonal antibody (mAb),1A2 and the highest titer was selected for further research. The mAb 1A2,an IgG1 type antibody was bonded to its immunizing antigen, both the eukaryotic and natural ORFV118 with high specificity. The immunohistochemical analysis showed that the focal specific staining was restricted to the epidermal layer and subcutaneous tissue, which conformed to the characteristics of an ORFV infection. The mAb 1A2 recognized ORFV118with high specificity. Further study of mAb 1A2 will facilitate our understanding of ORFV118 and provide potentially novel methods for the diagnosis, prevention, and treatment of Orf.

[Preparation and identification of rabbit polyclonal antibodies against vacuolar proton pyrophosphatase type I of Toxoplasma gondii].

OBJECTIVE: To prepare and characterize rabbit polyclonal antibodies against Toxoplasma gondii vacuolar proton pyrophosphatase type I (TgVP1). METHODS AND RESULTS: Two synthesized peptides TgVP1-1 and TgVP1-2 as the haptens were conjugated with KLH to immunize rabbits. Indirect ELISA showed that the titers of rabbit anti-TgVP1-1 polyclonal antibody and rabbit anti-TgVP1-2 polyclonal antibody reached 1:128 000. Western blotting results revealed that both purified polyclonal antibodies could specifically bind to a purified 85 kD T. gondii protein predicted as TgVP1. The protein detected by these two polyclonal antibodies was distributed in the cytoplasm of T. gondii tachyzoite, and this distribution pattern was consistent with that of acidocalcisome. CONCLUSION: The peptide-based method of antibody generation is efficient and the obtained TgVP1 polyclonal antibodies possess a high specificity to facilitate further study of T. gondii acidocalcisome and the diagnosis of toxoplasmosis.

Generation and Identification of Peptide-Based Monoclonal Antibodies Against Vacuolar Proton Pyrophosphatase of Toxoplasma gondii.

Vacuolar proton pyrophosphatase (V-PPase), an electrogenic proton pump widely distributed in non-mammalian species, is one of the important targets for acidocalcisomes. In this study, a novel method of peptide-based antibody generation was performed to produce monoclonal antibodies (MAbs) against Toxoplasma gondii V-PPase. Three hybridomas were identified and confirmed by ELISA, Western blotting, and immunofluorescence. All of them can react with an 85 kDa band of T. gondii protein in purified acidocalcisomal fraction. The three MAbs were all specific to the synthetic peptide of YTKAADVGADLSGKNEYGMSEDDPRNPAC, corresponding to amino acids at the location of 292aa-320aa of TgVP1 amino acid sequence. These specific MAbs will be valuable tools for further study of T. gondii infection biology, pathogenesis, and host immune response.

Development and calibration of a single UV LED based bioaerosol monitor.

A high-sensitivity bioaerosol monitor based on a single 365 nm LED is developed and a calibration approach is discussed for the first time. The fluorescence detection system, which is the core part of the monitor, contains an optical detection module and a fluorescence signal processor configured with a phase sensitive detector (PSD). B800 fluorescent microspheres and staphylococcus are used for performance evaluation of the monitor. B800 microspheres are appropriate as calibration material. The experimental results demonstrate the PSD plays a significant role in improving sensitivity and signal-to-noise ratio (SNR) of detection. Our monitor can detect staphylococcus concentration above 1800 cfu/L of air stably.

Taqman real-time PCR assay based on ORFV024 gene for rapid detection of orf infection.

In this study, a TaqMan real-time polymerase chain reaction (PCR) assay was developed to detect and quantify orf virus (ORFV) DNA in infected cell culture and clinical samples. Primers and probes were designed to amplify an 87 bp fragment DNA based on the sequence of ORFV024 gene encoding an NF-κB inhibitor of orf virus. The assay was highly specific and sensitive for ORFV DNA and no cross-reactions were detected with any other poxviruses; the sensitivity was 5 fg or 15 copies of ORFV genomic DNA. Both intra- (1.490 ± 1.261%) and inter-assay (1.958 ± 0.568%) variabilities were within the acceptable range, indicating the high efficiency and reproducibility of the assay. Further, the assay has shown a relative diagnostic sensitivity and specificity of 100%, when compared to B2L gene-based semi-nested PCR. The assay is simple, rapid, specific and sensitive with a wide potential for rapid field diagnosis of orf in sheep and goats.

Isolation and phylogenetic analysis of orf virus from the sheep herd outbreak in northeast China.

BACKGROUND: Orf is a zoonotic and epitheliotrophic contagious disease that mainly affects sheep, goats, wild ruminants, and humans with a worldwide distribution. To date, there is little information on the characterization of ORFV strains that are endemic in Mainland China. In addition, the relationship between the severity of disease and the molecular profile of ORFV strains has not been fully elucidated. RESULTS: From the recent outbreak of a sheep herd in Nongan, northeast of China, the novel orf virus (ORFV) strain NA1/11 was successfully isolated. Western blot analysis indicated that the NA1/11 strain cross reacts with monoclonal antibody A3 and infected sheep ORFV antiserum. The purified virions revealed the typical ovoid shape when observed by atomic force microscopy. To determine the genetic characteristics of the NA1/11 strain, the sequences of ORFV011 (B2L), ORFV059 (F1L), ORFV109, ORFV110 and ORFv132 (VEGF) genes were amplified and compared with reference parapoxvirus strains. Non-metric multidimensional scaling (nMDS) was performed to analyze the nucleotide similarities between different ORFV strains. CONCLUSIONS: Phylogenetic analysis based on ORFV 011 nucleotide sequences showed that the NA1/11strain was closely related to Xinjiang and Gansu strains. ORFV110 and ORFV132 genes are highly variable. The results revealed that precise phylogenetic analysis might provide evidence for genetic variation and movement of circulating ORFV strains in Northeast China. In addition, nMDS analysis showed that geographic isolation and animal host are likely major factors resulting in genetic differences between ORFV strains.

Identification and characterization of monoclonal antibodies against the ORFV059 protein encoded by Orf virus.

Recent outbreaks of orf in China have been attributed to a novel strain of Orf virus (ORFV) designated ORFV-Jilin. Currently, monoclonal antibodies (Mabs) have not yet been developed against this specific pathogen even though such entities could have potential applications regarding the diagnosis and characterization of ORFV-Jilin. Therefore, the current study was undertaken to generate Mab against the immunodominant ORFV059 protein of this virus. For this purpose, the ORFV-Jilin ORFV059 protein was expressed in Escherichia coli and subsequently used as an antigen to immunize mice and for the initial screening of hybridomas prepared from the mice for their ability to produce anti-ORFV059 protein Mabs via an indirect ELISA. Ten, positive hybridomas were identified in this manner and verified based on the ability of their released Mab to react specifically with both naturally and artificially expressed ORFV059 protein in Western blots. The two hybridomas with the greatest propensity to secrete Mab were subcloned three times before being introduced intraperitoneally into mice. Afterwards, both Mab were separately purified from the mice's ascetic fluids and found to successfully recognize the ORFV-Jilin ORFV059 protein in a variety of immunological assays. Thus, the widespread utility of these Mab as a diagnostic core reagent should prove invaluable for further investigations regarding the mechanisms of orf pathogenesis and the control of this disease. In this regard, it should be noted that Mab A3 was used to confirm the predicted late expression of the ORFV-Jilin ORFV059 protein during virus replication.