Plastid phylogenomics contributes to the taxonomic revision of taxa within the genus Sanicula L. and acceptance of two new members of the genus.

Introduction: The genus Sanicula L. is a taxonomically complicated taxa within Apiaceae, as its high variability in morphology. Although taxonomists have performed several taxonomic revisions for this genus, the interspecific relationships and species boundaries have not been satisfactorily resolved, especially for those endemic to China. This study mainly focused on S. giraldii var. ovicalycina, S. tienmuensis var. pauciflora, and S. orthacantha var. stolonifera and also described two new members of the genus. Methods: We newly sequenced sixteen plastomes from nine Sanicula species. Combined with eleven plastomes previously reported by us and one plastome downloaded, we performed a comprehensively plastid phylogenomics analysis of 21 Sanicula taxa. Results and Discussion: The comparative results showed that 21 Sanicula plastomes in their structure and features were highly conserved and further justified that two new species were indeed members of Sanicula. Nevertheless, eleven mutation hotspot regions were still identified. Phylogenetic analyses based on plastome data and the ITS sequences strongly supported that these three varieties were clearly distant from three type varieties. The results implied that these three varieties should be considered as three independent species, which were further justified by their multiple morphological characters. Therefore, revising these three varieties into three independent species was reasonable and convincing. Moreover, we also identified and described two new Sanicula species (S. hanyuanensis and S. langaoensis) from Sichuan and Shanxi, China, respectively. Based on their distinct morphological characteristics and molecular phylogenetic analysis, two new species were included in Sanicula. In summary, our study impelled the revisions of Sanicula members and improved the taxonomic system of the genus.

POU2F2-mediated upregulation of lncRNA PTPRG-AS1 inhibits ferroptosis in breast cancer via miR-376c-3p/SLC7A11 axis.

Background: Triple-negative breast cancer (TNBC) is a subtype of BC with high rates of mortality. The mechanism of PTPRG-AS1 in ferroptosis of TNBC was investigated. Methods: Chromatin immunoprecipitation and dual-luciferase reporter assays were used to measure intermolecular relationships. MTT and colony formation assays detected cell viability and proliferation. Kits detected Fe2+ and reactive oxygen species levels. The role of PTPRG-AS1 in tumor growth was analyzed in vivo. Results: PTPRG-AS1 was increased in TNBC tissues and cells. PTPRG-AS1 silencing increased the reduction of glutathione and GPX4, increased Fe2+ and reactive oxygen species in erastin-treated cells and inhibited proliferation. POU2F2 transcriptionally upregulated PTPRG-AS1. PTPRG-AS1 targeted miR-376c-3p to upregulate SLC7A11. PTPRG-AS1 knockdown suppressed tumor growth in vivo. Conclusion: POU2F2 transcriptionally activates PTPRG-AS1 to modulate ferroptosis and proliferation by miR-376c-3p/SLC7A11, promoting TNBC.

Triple-negative breast cancer (TNBC) is a kind of breast cancer with high recurrence and low survival rates. Activation of the ferroptosis pathway can inhibit BC proliferation and distant metastasis. Therefore, identifying effective biomarkers and molecular mechanisms of ferroptosis in TNBC is important for its earlier detection and therapy. PTPRG-AS1 is a new type of lncRNA discovered in recent years that is increased in various diseases and is related to prognosis. In the present study, the authors found that POU2F2 promoted PTPRG-AS1 transcription. PTPRG-AS1 knockdown activated ferroptosis in TNBC and inhibited proliferation. Mechanistically, PTPRG-AS1 targeted miR-376c-3p to upregulate SLC7A11, thereby inhibiting ferroptosis and promoting TNBC development. These results indicate that PTPRG-AS1 is a possible therapeutic target in TNBC.

Phylogeny and diversification of genus Sanicula L. (Apiaceae): novel insights from plastid phylogenomic analyses.

BACKGROUND: The genus Sanicula L. is a unique perennial herb that holds important medicinal values. Although the previous studies on Sanicula provided us with a good research basis, its taxonomic system and interspecific relationships have not been satisfactorily resolved, especially for those endemic to China. Moreover, the evolutionary history of this genus also remains inadequately understood. The plastid genomes possessing highly conserved structure and limited evolutionary rate have proved to be an effective tool for studying plant phylogeny and evolution. RESULTS: In the current study, we newly sequenced and assembled fifteen Sanicula complete plastomes. Combined with two previously reported plastomes, we performed comprehensively plastid phylogenomics analyses to gain novel insights into the evolutionary history of this genus. The comparative results indicated that the seventeen plastomes exhibited a high degree of conservation and similarity in terms of their structure, size, GC content, gene order, IR borders, codon bias patterns and SSRs profiles. Such as all of them displayed a typical quadripartite structure, including a large single copy region (LSC: 85,074-86,197 bp), a small single copy region (SSC: 17,047-17,132 bp) separated by a pair of inverted repeat regions (IRs: 26,176-26,334 bp). And the seventeen plastomes had similar IR boundaries and the adjacent genes were identical. The rps19 gene was located at the junction of the LSC/IRa, the IRa/SSC junction region was located between the trnN gene and ndhF gene, the ycf1 gene appeared in the SSC/IRb junction and the IRb/LSC boundary was located between rpl12 gene and trnH gene. Twelve specific mutation hotspots (atpF, cemA, accD, rpl22, rbcL, matK, ycf1, trnH-psbA, ycf4-cemA, rbcL-accD, trnE-trnT and trnG-trnR) were identified that can serve as potential DNA barcodes for species identification within the genus Sanicula. Furthermore, the plastomes data and Internal Transcribed Spacer (ITS) sequences were performed to reconstruct the phylogeny of Sanicula. Although the tree topologies of them were incongruent, both provided strong evidence supporting the monophyly of Saniculoideae and Apioideae. In addition, the sister groups between Saniculoideae and Apioideae were strongly suggested. The Sanicula species involved in this study were clustered into a clade, and the Eryngium species were also clustered together. However, it was clearly observed that the sections of Sanicula involved in the current study were not respectively recovered as monophyletic group. Molecular dating analysis explored that the origin of this genus was occurred during the late Eocene period, approximately 37.84 Ma (95% HPD: 20.33-52.21 Ma) years ago and the diversification of the genus was occurred in early Miocene 18.38 Ma (95% HPD: 10.68-25.28 Ma). CONCLUSION: The plastome-based tree and ITS-based tree generated incongruences, which may be attributed to the event of hybridization/introgression, incomplete lineage sorting (ILS) and chloroplast capture. Our study highlighted the power of plastome data to significantly improve the phylogenetic supports and resolutions, and to efficiently explore the evolutionary history of this genus. Molecular dating analysis explored that the diversification of the genus occurred in the early Miocene, which was largely influenced by the prevalence of the East Asian monsoon and the uplift of the Hengduan Mountains (HDM). In summary, our study provides novel insights into the plastome evolution, phylogenetic relationships, taxonomic framework and evolution of genus Sanicula.

Crosstalk Between Peripheral Innervation and Pancreatic Ductal Adenocarcinoma / 神经科学通报·英文版

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive lethal malignancy, characterized by late diagnosis, aggressive growth, and therapy resistance, leading to a poor overall prognosis. Emerging evidence shows that the peripheral nerve is an important non-tumor component in the tumor microenvironment that regulates tumor growth and immune escape. The crosstalk between the neuronal system and PDAC has become a hot research topic that may provide novel mechanisms underlying tumor progression and further uncover promising therapeutic targets. In this review, we highlight the mechanisms of perineural invasion and the role of various types of tumor innervation in the progression of PDAC, summarize the potential signaling pathways modulating the neuronal-cancer interaction, and discuss the current and future therapeutic possibilities for this condition.

Enzalutamide and olaparib synergistically suppress castration-resistant prostate cancer progression by promoting apoptosis through inhibiting nonhomologous end joining pathway / 亚洲男科学杂志(英文版)

Recent studies revealed the relationship among homologous recombination repair (HRR), androgen receptor (AR), and poly(adenosine diphosphate-ribose) polymerase (PARP); however, the synergy between anti-androgen enzalutamide (ENZ) and PARP inhibitor olaparib (OLA) remains unclear. Here, we showed that the synergistic effect of ENZ and OLA significantly reduced proliferation and induced apoptosis in AR-positive prostate cancer cell lines. Next-generation sequencing followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed the significant effects of ENZ plus OLA on nonhomologous end joining (NHEJ) and apoptosis pathways. ENZ combined with OLA synergistically inhibited the NHEJ pathway by repressing DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and X-ray repair cross complementing 4 (XRCC4). Moreover, our data showed that ENZ could enhance the response of prostate cancer cells to the combination therapy by reversing the anti-apoptotic effect of OLA through the downregulation of anti-apoptotic gene insulin-like growth factor 1 receptor ( IGF1R ) and the upregulation of pro-apoptotic gene death-associated protein kinase 1 ( DAPK1 ). Collectively, our results suggested that ENZ combined with OLA can promote prostate cancer cell apoptosis by multiple pathways other than inducing HRR defects, providing evidence for the combined use of ENZ and OLA in prostate cancer regardless of HRR gene mutation status.

Plastid phylogenomic insights into relationships, divergence, and evolution of Apiales.

MAIN CONCLUSION: Members of Apiales are monophyletic and radiated in the Late Cretaceous. Fruit morphologies are critical for Apiales evolution and negative selection and mutation pressure play important roles in environmental adaptation. Apiales include many foods, spices, medicinal, and ornamental plants, but the phylogenetic relationships, origin and divergence, and adaptive evolution remain poorly understood. Here, we reconstructed Apiales phylogeny based on 72 plastid genes from 280 species plastid genomes representing six of seven families of this order. Highly supported phylogenetic relationships were detected, which revealed that each family of Apiales is monophyletic and confirmed that Pennanticeae is a member of Apiales. Genera Centella and Dickinsia are members of Apiaceae, and the genus Hydrocotyle previously classified into Apiaceae is confirmed to belong to Araliaceae. Besides, coalescent phylogenetic analysis and gene trees cluster revealed ten genes that can be used for distinguishing species among families of Apiales. Molecular dating suggested that the Apiales originated during the mid-Cretaceous (109.51 Ma), with the families' radiation occurring in the Late Cretaceous. Apiaceae species exhibit higher differentiation compared to other families. Ancestral trait reconstruction suggested that fruit morphological evolution may be related to shifts in plant types (herbaceous or woody), which in turn is related to the distribution areas and species numbers. Codon bias and positive selection analyses suggest that negative selection and mutation pressure may play important roles in environmental adaptation of Apiales members. Our results improve the phylogenetic framework of Apiales and provide insights into the origin, divergence, and adaptive evolution of this order and its members.

Potential Chemoprevention of Paeoniflorin in Colitis-Associated Colorectal Cancer by Network Pharmacology, Molecular Docking, and In Vivo Experiment.

Chronic inflammation plays a positive role in the development and progression of colitis-associated colorectal cancer (CAC). Medicinal plants and their extracts with anti-inflammatory and immunoregulatory properties may be an effective treatment and prevention strategy for CAC. This research aimed to explore the potential chemoprevention of paeoniflorin (PF) for CAC by network pharmacology, molecular docking technology, and in vivo experiments. The results showed that interleukin-6 (IL-6) is a key target of PF against CAC. In the CAC mouse model, PF increased the survival rate of mice and decreased the number and size of colon tumors. Moreover, reduced histological score of colitis and expression of Ki-67 and PCNA were observed in PF-treated mice. In addition, the chemoprevention mechanisms of PF in CAC may be associated with suppression of the IL-6/STAT3 signaling pathway and the IL-17 level. This research provides experimental evidence of potential chemoprevention strategies for CAC treatment.

Barriers and facilitators of lung cancer screening uptake: protocol of a mixed methods systematic review.

INTRODUCTION: The global uptake rates of lung cancer screening (LCS) with low-dose CT remain low. Since numerous factors contribute to the underuse of LCS, a theory-informed approach to identify and address the uptake of LCS barriers and facilitators is required. This study aims to document the methods which were used to identify, appraise, and synthesise the available qualitative, quantitative, and mixed methods evidence, addressing the barriers and facilitators at the individual and healthcare provider level, according to the social-ecological model, before identifying gaps to aid future practices and policies. METHODS AND ANALYSIS: The following databases will be searched: PubMed, Ovid (Journals @ Ovid Full Text and Ovid MEDLINE), EMBASE, CINAHL, PsycINFO, Cochrane Library, Chinese Biomedical Database, Chinese National Knowledge Infrastructure, and Wanfang database, from their creation up to 31 December 2020. Two reviewers will be involved in independently screening, reviewing, and synthesising the data; and calibration exercises will be conducted at each stage. Disagreements between the two reviewers will be resolved by arbitration by a third reviewer. The Critical Appraisal Checklist for Studies Reporting Prevalence Data from the Joanna Briggs Institute, the Critical Appraisal Skills Programme criteria adapted for qualitative studies, and the 16-item Quality Assessment Tool (QATSDD) will be used in the quality assessment of primary studies. We will perform data synthesis using the Review Manager software, V.5.3. ETHICS AND DISSEMINATION: This study is a review of published data and therefore needs no ethical approval. The findings of this systematic review will be published in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: CRD42020162802.

Identification and inner relation between gastrointestinal stromal tumor and intra-abdominal desmoid tumor / 中华胃肠外科杂志

Intra-abdominal desmoid tumor (IADT) and gastrointestinal stromal tumor (GIST) are both mesenchymal tumors mostly found in gastrointestinal tracts and easily misdiagnosed, which would directly damage the survival prognosis and quality of life of patients. With the advent of the era of precision medicine, the understanding of the above two diseases is more in-depth, and the requirements for accurate diagnosis and individualized precision treatment are more stringent. Moreover, there seems to be some internal relationship between IADT and GIST, and the lack of systematic research and discussion makes clinical decision-making and patient management easy to fall into traps and misunderstandings. Therefore, this paper reviews the clinical characteristics, pathogenesis and treatments of the two, and explore their differences and internal relations, so as to provide research and practical reference for promoting more precise and individualized diagnosis and treatment regimens.

Research progress of long bone dysplasia caused by glucocorticoid exposure during pregnancy / 中国药理学通报

The main form of fetal long bone development is endochondral ossification. In recent years, studies have shown that the mother during pregnancy with bad environmental exposure to high levels of endogenous glucocorticoid (GC) and premature use of artificial synthetic GC can go through the placenta into the fetal body, cause fetal blood GC levels, leading to intrauterine retardation, and affect fetal cartilage ossification. This effect can extend beyond birth and even into old age, leading to susceptibility and heritability of osteoporosis in offsprings. This review summarizes the current status of glucocorticoid exposure during pregnancy, summarizes the short-term and long-term effects of intrauterine GC exposure on long bone development in offsprings, and explains the possible mechanism of intrauterine endocrine programming, which lays a theoretical foundation for the prevention and treatment of fetal bone diseases caused by GC exposure during pregnancy and the future research direction of developmental diseases.

UCH-L1-mediated Down-regulation of Estrogen Receptor α Contributes to Insensitivity to Endocrine Therapy for Breast Cancer.

Purpose: To determine the role of UCH-L1 in regulating ERα expression, and to evaluate whether therapeutic targeting of UCH-L1 can enhance the efficacy of anti-estrogen therapy against breast cancer with loss or reduction of ERα. Methods: Expressions of UCH-L1 and ERα were examined in breast cancer cells and patient specimens. The associations between UCH-L1 and ERα, therapeutic response and prognosis in breast cancer patients were analyzed using multiple databases. The molecular pathways by which UCH-L1 regulates ERα were analyzed using immunoblotting, qRT-PCR, immunoprecipitation, ubiquitination, luciferase and ChIP assays. The effects of UCH-L1 inhibition on the efficacy of tamoxifen in ERα (-) breast cancer cells were tested both in vivo and in vitro. Results: UCH-L1 expression was conversely correlated with ERα status in breast cancer, and the negative regulatory effect of UCH-L1 on ERα was mediated by the deubiquitinase-mediated stability of EGFR, which suppresses ERα transcription. High expression of UCH-L1 was associated with poor therapeutic response and prognosis in patients with breast cancer. Up-regulation of ERα caused by UCH-L1 inhibition could significantly enhance the efficacy of tamoxifen and fulvestrant in ERα (-) breast cancer both in vivo and in vitro. Conclusions: Our results reveal an important role of UCH-L1 in modulating ERα status and demonstrate the involvement of UCH-L1-EGFR signaling pathway, suggesting that UCH-L1 may serve as a novel adjuvant target for treatment of hormone therapy-insensitive breast cancers. Targeting UCH-L1 to sensitize ER negative breast cancer to anti-estrogen therapy might represent a new therapeutic strategy that warrants further exploration.

Retrospective analysis of the on-site treatment of the coronavirus disease 2019 epidemic on the Costa Crociere cruise / 中华危重病急救医学

Coronavirus disease 2019 (COVID-19) epidemic is the most widespread global pandemic in the past 100 years. Person-to-person transmission of COVID-19 infection leads to the major threat of human safety and health. At 00:00 on January 24th, 2020, Tianjin City launched the first-level response to the COVID-19 epidemic. At 18:00 on the same day, Management Committee of Dongjiang Free Trade Port Zone of Tianjin received areport that there were 15 people who had fever on the Costa Crociere carrying 4 806 people from Japan back to the home port of Tianjin Dongjiang Cruise. At the same time, there are more than 140 Chinese Hubei tourists. Tianjin Municipal Committee and Government, Tianjin Customs, Binhai New Area District Committee Government, Tianjin Health Commission, Tianjin Binhai New Area Health Commission formed an emergency command center immediately to deal with the epidemic comprehensively. At 06:40 on January 25th, 2020, the medical investigation team made up by Tianjin Binhai New Area Health Commission and Tianjin East Administration of Customs boarded the cruise ship. With reference to the customs inspection and quarantine regulations, in accordance with the Diagnosis and treatment of pneumonia caused by novel coronavirus (trial version 3) for mulated by the National Health Commission of the People's Republic of China and the Novel coronavirus infected pneumonia port control and technology plan (first version) formulated by the General Administration of Customs, combined with the actual situation of cruise ships, the medical investigation team developed the inspection standards, including door-to-door inspections, temperature measurement and epidemiological investigations on all persons on board of the cruise ship. A total of 4 806 person-times were investigated in the affected area, including 3 706 tourists and 1 100 crew members. Seventeen people at high risk of COVID-19 were identified, including three Wuhan tourists. The reports of 2019 novel coronavirus (2019-nCoV) nucleic acid detection on throat swab samples for those who were identified as high risk were returned as all negative at 14:54 on the same day. At 19:30, the medical investigation team completed the investigation and evacuated the cruise ship. The temperature measurement, medical observation and resettlement of passenger were handed over to relevant personnel. After 2 weeks, the follow-up result of 2019-nCoV nucleic acid of 17 high risk people were all negative. The overall command and comprehensive coordination of the onshore command center together with the rigid principles and excellent responds ability of the on-site epidemic investigation team ensured the successful completion of the epidemic investigation work, and also provided reference for further improving the management and disposal capacity of public health emergencies at sea.

Analysis of the associations between anxiety and different family rearing styles and sleep habits in preschool children / 中国医师进修杂志

Objective:To explore the associations between anxiety and family rearing styles and sleep habits in preschool children.Methods:From March to October 2019, a total of 100 preschool children (47 boys and 53 girls) were investigated by Spence Children′s anxiety scale, Family rearing style Evaluation scale (parents′ version) and self-made sleep habit questionnaire.Results:There was no significant difference in the total score of anxiety between girls and boys: (55.12 ± 9.89) scores vs. (53.40 ± 11.82) scores, P > 0.05. The parent′s acrasia-control was positively correlated with children′s social anxiety dimension and total score of anxiety ( r = 0.202 and 0.202, P<0.05), the parent′s reception-rejection was positively correlated with children′s generalized anxiety dimension ( r = 0.237, P<0.05), and children′s sleep duration was negatively correlated with physical injury fear dimension and the total score of anxiety ( r = -0.230 and -0.203, P<0.05). The anxiety scores in children who slept in the same bed was significantly higher than that in children who slept alone and who slept in different beds in the same room: (53.57 ± 9.75) scores vs. (50.21 ± 13.89) and (50.48 ± 11.50) scores, and there was statistical difference ( P<0.05); there was no difference in the total score of anxiety between children who slept alone and who slept in different beds ( P>0.05). Multivariate analysis result showed that children′s age, sex, sleep duration, parents′ acrasia-control and partial-rough were the predictors of children′s anxiety ( P<0.05 or<0.01). Conclusions:Different family rearing styles and children′s bed sleeping styles significantly affect the anxiety of preschool children. Parents should pay attention to the positive effects of good rearing styles and children′s separate sleep on anxiety.

Collaborative study to evaluate a reporter gene assay for anti-PD-1 antibody bioactivity / 药学学报

A collaborative inter-laboratory validation was carried out using a reporter gene assay to measure the bioactivity of anti-PD-1 monoclonal antibody, in order to study the applicability and transferability of the method. In this study, two collaborative schemes were designed to measure the precision, linearity and accuracy of the method. The results showed that the 95% confidence interval (CI) of the intra-assay precision was (1.72-16.89) %, inter-assay precision was (2.63-17.67) %, inter-laboratory precision was (9.00-14.26) %, all linear correlation coefficients were greater than 0.99, and the 95% CI for the accuracy at different potency levels was (91.83-104.40) % at 50%, (90.40-101.40) % at 75%, (94.71-105.60) % at 100%, (94.00-102.00) % at 125%, and (96.73-104.30) % at 150%. The collaborative validation results proved that the reporter gene assay for the bioactivity determination of anti-PD-1 monoclonal antibody has good precision, linearity and accuracy, and could be applied to the release and stability analysis of anti-PD-1 monoclonal antibodies in different laboratories.

Effect of triglyceride level on acute kidney injury complicating acute pancreatitis / 上海预防医学

[Objective] To explore whether the triglyceride level is associated with incidence of acute kidney injury in patients with acute pancreatitis. [Methods] From Jan 2015 to March 2017, 184 patients with acute pancreatitis were selected and divided in to 3 groups based on different triglyceride levels: ideal triglyceride group (n = 89) , mild high triglyceride group (n = 53) and severe high triglyceride group (n =42). The incidence of acute kidney injury and its severity were compared between the three groups.[Results]The incidence of acute kidney injury in severe high triglyceride group was 33.3％, significantly higher than that in ideal triglyceride group (12.3％). The surgical treatment (16.7％ vs 4.5％) , average hospitalization days (20 days vs 14 days) and systemic inflammatory response syndrome score (3.0 vs2.3) in severe high triglyceride group were higher than those in ideal triglyceride group, and all differences between the two groups were significant. After adjusting for factors such as age, sex, body mass index and other confounders, the risk of acute kidney injury occurring in severe high triglyceride group was 2.35 times that in ideal triglyceride group (95％CI: 1.32-4.29). [Conclusion] High triglyceride level proves to be associated with high risk of acute kidney injury in patients with acute pancreatitis.

Liuwei Dihuang Pill () Treats Postmenopausal Osteoporosis with Shen (Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression / 中国结合医学杂志

<p><b>OBJECTIVES</b>To investigate the mechanism of Liuwei Dihuang Pill (, LDP) in treating postmenopausal osteoporosis (PMOP) with Shen (Kidney) yin deficiency.</p><p><b>METHODS</b>In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group (Group A), PMOP Shen-yang deficiency group (Group B), PMOP without Shen deficiency group (Group C), and control group (Group N). Real-time polymerase chain reaction (RT-PCR) and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 (CLCF1), ankyrin repeat and SOCS box containing 1 (ASB1), and prokineticin 2 (PROK2) genes and the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway.</p><p><b>RESULTS</b>The mRNA (P<0.05) and protein (P<0.01) expression levels of the CLCF1 gene in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gene were obviously up-regulated (P<0.01). After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated (both P<0.01), and the average bone density of the top femur had significantly increased (P<0.05). In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3.</p><p><b>CONCLUSIONS</b>The CLCF1 gene is an important gene associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gene expression and activation of the JAK/STAT signaling pathway.</p>

Application of native hapten-gel diffusion test in differential diagnosis of Brucella-infected antibody and S2 or Rev.1-vaccinated antibody / 中国人兽共患病学报

The aim of the present study was to work on the efficiency of differential diagnosis of native hapten-gel diffusion assay (NH-GD) on the background of vaccination with S2 or Rev.1.The conditions of NH-GD assay was firstly optimized,its sensitivity,specificity,repeatability and ability of differential diagnosis were determined respectively,and its test result was compared with that of fluorescence polarization assay (FPA).The results showed NH-GD assay with good specificity and repeatability could differentiate Brucella-infected antibody from vaccinated antibody after vaccination with S2 or 122 days after vaccination with Rev.1.And the result of NH-GD assay was highly consistent with that of FPA,which was simple to operate and needed a few simple equipment.Therefore,NH-GD assay was a good method for sheep brucellosis surveillance in China and especially suitable for application in grass-roots areas.

Specific sIgA antibodies against swine influenza virus H3N2 infection / 中国人兽共患病学报

To study different breed pigs reply the swine flu virus infections,specific antibody of sIgA secretion regularity of respiratory tract and the differences of sIgA antibody according to different antigen proteins were detected.A/swine/Nanjing/ 51/2010(H3N2) was intranasally infected pigs (1 × 107 TCID50/mL and 2 mL/pig),and then the nasal swab samples were collected at different time points within 21 days after infection.M1,NS1 and PB1 recombinant protein,respectively,were used to establish indirect ELISA method for detecting specific antibody of sIgA,and to analyze its secretion regularity.Results displayed that there was no significant difference among three kinds of recombinant protein in the whole test,characterizing by specificity sIgA antibody levels rising rapidly after 5 infection days and reaching peak at day 14,then began to decline.Among different varieties of pigs,sIgA antibody production of PB1 protein in Obama group was significantly higher than that in binary pigs at 14th and 21st day (P＜0.05).It had no significant difference between M1 group and the NS1 group (P＞0.05).This experiment preliminary explores the secretion regularity of specificity sIgA antibody after infected swine flu virus,which laid a foundation for further study of SIV mucosal antibody diagnostic reagents.

Preparation and identification of a monoclonal antibody against Mycoplasma hyorhinis / 中国人兽共患病学报

The aim of this study was to produce a monoclonal antibody against Mycoplasma hyorhinis,which would be useful in the diagnosis and pathogenesis researches of M.hyorhinis.BALB/c mice were immunized with whole cell protein of M.hyorhinis,and then the monoclonal antibody (McAb) was prepared by hybridoma technique.The isotype of the McAb was identified,and then the titer and specificity were analyzed.The prepared McAb was used to detect M.hyorhinis in the colony immunoboltting assay and indirect immunofluorescence assay.A hybridoma cell line was obtained,and was named Mhr-08.The McAb belonged to IgG1 isotype,and the light chain was κ type.The titer was detected to be 1 ∶ 102 400 by ELISA.The result of Western-blot indicated that this McAb strongly reacted to a 43 kDa protein of M.hyorhinis,without cross-reaction with other swine mycoplasmas,Escherichia coli or KM2 medium.It revealed that the McAb recognized a surface membrane protein and was successfully applied to detect M.hyorhinis in colony immunoboltting assay.The mycoplasmas bound to the tracheal epithelial cells was successfully detected by using this McAb in indirect immunofluorescence assay.In conclusion,a McAb against M.hyorhinis was successfully prepared,which provides as a useful tool for the diagnosis and pathogenesis researches.

[Effect of ethanol and its metabolites on acetylcholine-sensitive K(+) channel Kir3.1 protein expression of neonatal rat primary atrial cardiomyocytes].

OBJECTIVE: To identify the effect of ethanol and its metabolite acetaldehyde on acetylcholine-sensitive K(+) channel Kir3.1 protein expression, and explore the potential role of this channel and acetaldehyde in arrhythmia caused by acute alcoholic intoxication. METHODS: Primary atrial cardiomyocytes were isolated from 150 newborn SD rats by typsin and type II collagenase, cultured and troponin I was determined by immunofluorescence. Cell survival in 200-800 mmol/L ethanol or 50-500 µmol/L acetaldehyde treated cells for 24 hours was measured by CCK-8 assay to determine the concentration of ethanol and acetaldehyde for inducing apoptosis in cardiomyocytes. The highest non-apoptotic concentration (200 mmol/L) of ethanol and acetaldehyde (100 µmol/L) was used in the main study. Kir3.1 protein expression was detected by Western blot. RESULTS: (1) Cellular immunofluorescence results showed that cultured cells are cardiomyocytes, and more than 90% of these cells are troponin I positive. (2) CCK-8 assay demonstrated that the survival rate of cardiomyocytes in the groups treated by ethanol over 400 mmol/L for 24 hours or acetaldehyde over 400 µmol/L was significantly lower than that of the control group (P < 0.05), while the survival rate was similar in cardiomyocytes treated by ethanol less than 200 mmol/L or acetaldehyde less than 350 µmol/L for 24 hours and the control group (P > 0.05). (3) Western-bolt assay revealed that ethanol and acetaldehyde treatment for 24 hours upregulated Kir3.1 protein expression in primary atrial cardiomyocytes of newborn SD rats by (44.52 ± 23.07)% and (45.04 ± 22.01)% respectively compared with the control group (all P < 0.01). CONCLUSIONS: Acute ethanol and acetaldehyde treatment could significantly upregulate the protein expression of acetylcholine-sensitive K(+) channel Kir3.1, this might serve as a potential mechanism for arrhythmia caused by acute alcoholic intoxication.