The association of visceral pleural invasion with skip N2 metastasis on clinical stage IA NSCLC

Abstract Background Lung lymphatic drainage occurs mainly through a peribronchial path, but it is hypothesized that visceral pleural invasion could alter this path. This study aims to investigate the association between visceral pleural invasion, node upstaging, and N2 skip metastasis and the impact on survival in a population of patients with non-small cell lung cancer of 3 cm or smaller. Methods We retrospectively queried our institutional database of lung cancer resection for all patients with clinical stage IA NSCLC between June 2009 and June 2022. We collected baseline characteristics and clinical and pathological staging data. Patients were classified into two groups: The non-VPI group with negative visceral pleural invasion and the VPI group with positive. The primary results analyzed were the occurrence of nodal upstaging, skip N2 metastasis and recurrence. Results There were 320 patients analyzed. 61.3 % were women; the median age was 65.4 years. The pleural invasion occurred in 44 patients (13.7 %). VPI group had larger nodules (2.3 vs. 1.7 cm; p < 0.0001), higher 18F-FDG uptake (7.4 vs. 3.4; p < 0.0001), and lymph-vascular invasion (35.7 % vs. 13.5 %, p = 0.001). Also, the VPI group had more nodal disease (25.6 % vs. 8.7 %; p = 0.001) and skip N2 metastasis (9.3 % vs. 1.8 %; p = 0.006). VPI was a statistically independent factor for skip N2 metastasis. Recurrence occurred in 17.2 % of the population. 5-year disease-free and overall survival were worse in the VPI group. Conclusions The visceral pleural invasion was an independent factor associated with N2 skip metastasis and had worse disease-free and overall survival.

The neuroprotective effects of Haikun Shenxi capsule medicated serum on N2a/APP695 cells based on PINKl-Parkin mitophagy pathway / 中国药理学通报

Aim To research the neuroprotective effect of Haikun Shenxi (HKSX) medicated serum on N2a/ App695 cells and the underlying mechanism. Methods HKSX medicated serum was prepared and carbohydrate components in it was analyzed using high performance thin layer chromatography (HPTLC) . N2a/ App695 cells were intervened with HKSX medicated serum, the cytotoxicity of HKSX medicated serum was measured by MTT; AP[_

Prevention and treatment of lipoic acid-niacin on blue-light-induced retinal damage in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the preventive effect and optimal drug dose of lipoic acid-niacin(N2L)against blue light-induced retinal damage in SD rats, and to explore its possible protective mechanism.METHODS: A total of 36 specific pathogen free-grade male SD rats of 150-200 g were selected and randomly divided into normal control group, blue light injury group, N2L low-dose group(1.0 mg/kg), N2L medium-dose group(2.5 mg/kg), N2L high-dose group(5.0 mg/kg), and physiological saline group, with 6 rats in each group. The normal control group was reared in a 12 h dark and light cycle, and the rest of the groups received 9 h of daily light exposure, 3 h of blue light irradiation with a wavelength of 455 nm and an intensity of 3000±50 lx, and 12 h of darkness to establish the injury model, and were exposed to light exposure for 14 d. For 14 consecutive durations, a 1 mL dose of the corresponding drug was injected intraperitoneally. The rats were reared for another 5 d with a regular 12 h light-dark cycle and were examined by electroretinography. Specimens were prepared by over anesthesia, HE staining, and the thickness of the outer nuclear layer was observed under a optical microscope; superoxide dismutases(SOD)activity was detected by CheKineTM SOD Activity Assay Kit; and the retinal Caspase-3, quinone oxidoreductase 1(NQO1), glutathione S transferase(GST), Bcl-2, and Bax protein expression in rat retina were detected by Western blot.RESULTS: The amplitude of b-wave in dark-vision ERG 3.0 and 10.0(cd·s)/m2 stimulated light, b-wave in bright-vision ERG 3.0(cd·s)/m2 stimulated light, and the amplitude of the 2nd wave peak of oscillatory potential were significantly lower in blue light injury group than that in the normal control group(all P&#x0026;#x003C;0.01), while the amplitude was significantly higher in the N2L medium-dose group than in the blue light injury group(all P&#x0026;#x003C;0.05), and was not statistically different from that of the normal control group; the thickness of the retina in the blue light injury group was decreased in the ONL compared with that of the normal control group(P&#x0026;#x003C;0.001), while in the N2L medium dose group, it was thicker than that of the blue light injury group(P&#x0026;#x003C;0.001), and there was no statistically significant difference from the normal control group; SOD activity was significantly higher in the N2L medium-dose group than in the remaining 5 groups(P&#x0026;#x003C;0.05); the expression of Caspase-3, Bax, and NQO1 in the blue light injury group was higher than that of the normal control group(all P&#x0026;#x003C;0.01), and expression of Bax and Caspase-3 was significantly lower in the N2L medium-dose group compared with the blue light injury group(all P&#x0026;#x003C;0.001), whereas GST, NQO1 and Bcl-2 were significantly increased(all P&#x0026;#x003C;0.01).CONCLUSION:A concentration of 2.5 mg/kg N2L can effectively antagonize the damaging effect of blue light on the retina of SD rats, and it is expected to be a preventive and curative drug for it.

Investigation on the antifungal activity of pyranium derivatives N2 / 药学实践杂志

Objective To study the antifungal activity of N2 derivatives. Methods The anti-fungal activity of N2 compounds was investigated by micro-liquid dilution. Then the activity of N2 compounds on hyphal and biofilm formation was investigated. Results N2 compounds had significant antifungal activity against Candida albicans. It also expressed actively inhibitory effect on hyphal and biofilm formation. The mechanism of its fungicidal function was to damage the structure of candida albicans’ cell membrane and cell wall. Conclusion The results showed that N2 had obvious antifungal activity against Candida albicans., which provided a new idea for the development of antifungal drugs and the solution of antifungal drugs resistance.

Research progress in antigenic evolution of influenza A (H3N2) virus / 中华微生物学和免疫学杂志

Influenza viruses are common pathogens causing respiratory infections in humans. Among the four seasonal influenza viruses, influenza A virus H3N2 has become the leading cause of seasonal influenza illness and death, posing a great threat to public health and the economy. Since it first emerged and caused a pandemic in 1968, H3N2 has been circulating repeatedly in human beings and continually evades host immune attack by antigenic drift, resulting in a decrease in vaccine efficacy. In this paper, the antigenic evolution of influenza A virus H3N2, the impact of antigenic evolution on the selection of vaccine strains and some models for predicting the evolution of influenza viruses were analyzed and reviewed, which paved the road for understanding the antigenic evolution of influenza virus and vaccine development.

The efficacy of postoperative radiotherapy in stage Ⅲ(N 2) non-small cell lung cancer: a meta analysis / 中华放射肿瘤学杂志

Objective:To conduct meta analysis to compare the effect of complete resection with or without postoperative radiotherapy (PORT) on survival in stage Ⅲ(N 2) non-small cell lung cancer (NSCLC). Methods:Relevant studies of the efficacy of PORT for stage Ⅲ(N 2) NSCLC were searched from Wanfang Data, PubMed, and Cochrane Library from January 2006 to January 2022. Literature screening, extraction of information and assessment of the risk of bias of the included literature was carried out by two independent researchers. Meta analysis was performed using R4.0.3 software. Results:A total of 12 publications consisting of 2992 patients were included, 1479 cases in the PORT group and 1513 cases in the control group. PORT improved the overall survival (OS) and disease free survival (DFS) compared to the control group. Fixed-effects model meta analysis of 6 randomized controlled trials showed that PORT did not significantly reduce the risk of death ( HR=0.98, 95% CI: 0.80-1.20). Fixed-effects model meta analysis of 6 retrospective studies showed that PORT improved prognosis ( HR=0.68, 95% CI: 0.59-0.79). PORT could improve OS of patients with multiple (station) metastasis of ipsilateral mediastinum and / or submandibular lymph nodes ( HR=0.89, 95% CI: 0.80-0.99). Conclusions:PORT could improve OS and DFS in stage Ⅲ(N 2) NSCLC. A trend towards benefit can be observed in the subgroup with multiple/multi-station N2 metastasis.

Genetic variation characteristics of HA gene of influenza A virus (H3N2) in Guizhou province from 2017 to 2019 / 中国基层医药

Objective:To analyze the genetic variation characteristics of the HA gene of influenza A virus (H3N2) in Guizhou province from 2017 to 2019. Methods:Twenty strains of influenza A virus (H3N2) were randomly selected from 10 network laboratories in Guizhou province for RNA extraction. Reverse transcriptase-polymerase chain reaction and sequencing were performed. The products were analyzed using bioinformatics software.Results:The nucleotide homology of the HA gene of the 20 strains was 97.7%-100%, which was highly homologous to the vaccine strains A/Hong-Kong/4801/2014 recommended by WHO in 2017 and A/Singapore-INFIMH/16-0019/2016 recommended by WHO in 2018, but they were significantly different from the vaccine strain A/Kansas/14/2017 recommended by WHO in 2019. Genetic analysis showed that the 20 strains were divided into two branches, and the strains that were prevalent in 2019 were located in different branches, with marked genetic differences. Key site analysis showed mutations in antigenic determinants A, B, C, and E and mutations in the anterior and posterior walls of receptor binding sites. Key site analysis also showed that there was an increase in the number of glycosylation sites compared with the vaccine strains prevalent in the same year. Genetic distance, antigen sites, and glycosylation sites were slightly different between virus strains prevalent in 2017-2018 and virus strains prevalent in 2019. Conclusion:The HA gene of the influenza A virus subtype H3N2 in Guizhou province from 2017 to 2019 showed heterogeneity and gene mutation, especially in 2019. Therefore, close monitoring of the genetic evolution of the influenza A virus subtype H3N2 is necessary.

Influenza A(H3N2) infection followed by separate COVID-19 infection / Infección por el virus de la gripe A (H3N2) seguida de infección separada por el virus causante de la COVID-19 / Infecção por influenza A (H3N2) seguida de infecção independente por COVID-19

ABSTRACT This study describes the case of a health professional infected first by influenza virus A(H3N2) and then by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 11 days later. Respiratory samples and clinical data were collected from the patient and from close contacts. RNA was extracted from samples and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to investigate the viruses. The patient presented with two different illness events: the first was characterized by fever, chest and body pain, prostration and tiredness, which ceased on the ninth day; RT-qPCR was positive only for influenza virus A(H3N2). Eleven days after onset of the first symptoms, the patient presented with sore throat, nasal congestion, coryza, nasal itching, sneezing and coughing, and a second RT-qPCR test was positive only for SARS-CoV-2; in the second event, symptoms lasted for 11 days. SARS-CoV-2 sequencing identified the Omicron BA.1 lineage. Of the patient's contacts, one was coinfected with influenza A(H3N2) and SARS-CoV-2 lineage BA.1.15 and the other two were infected only with SARS-CoV-2, one also with Omicron BA.1.15 and the other with BA.1.1. Our findings reinforce the importance of testing for different viruses in cases of suspected respiratory viral infection during routine epidemiological surveillance because common clinical manifestations of COVID-19 mimic those of other viruses, such as influenza.

RESUMEN Este estudio describe el caso de un profesional de la salud que contrajo la infección primero por el virus de la gripe A (H3N2) y a continuación por el coronavirus 2 del síndrome respiratorio agudo grave (SARS-CoV-2) 11 días después. Se recogieron muestras respiratorias y datos clínicos del paciente y sus contactos cercanos. Se extrajo ARN de muestras y se utilizó la reacción en cadena de la polimerasa cuantitativa con transcripción inversa (RT-qPCR, por su sigla en inglés) para investigar los virus. El paciente presentó dos procesos infecciosos distintos: el primero se caracterizó por fiebre, dolor corporal y torácico, postración y cansancio, que cesó en el noveno día. La prueba mediante RT-qPCR solo fue positiva en el virus de la gripe A (H3N2). Once días después del inicio de los primeros síntomas, el paciente manifestó dolor de garganta, congestión nasal, catarro, picazón nasal, estornudos y tos. Una segunda prueba mediante RT-qPCR solo fue positiva para el SARS-CoV-2 y durante este segundo proceso los síntomas duraron 11 días. La secuenciación del SARS-CoV-2 identificó el linaje ómicron BA.1. De los contactos del paciente, uno presentaba una coinfección por el virus de la gripe A (H3N2) y el linaje BA.1.15 del SARS-COV-2, y los otros dos presentaban infecciones únicamente por SARS-CoV-2, uno también del linaje ómicron BA.1.15 y el otro de BA.1.1. Estos hallazgos refuerzan la importancia de realizar pruebas para detectar diferentes virus en casos de sospecha de infección viral respiratoria durante la vigilancia epidemiológica de rutina porque las manifestaciones clínicas comunes de COVID-19 son similares a las de otros virus, como en el caso de la gripe.

RESUMO Este estudo descreve o caso de uma profissional de saúde infectada primeiro pelo vírus influenza A (H3N2) e, 11 dias depois, pelo coronavírus da síndrome respiratória aguda grave 2 (SARS-CoV-2). Amostras respiratórias e dados clínicos foram coletados da paciente e de contatos próximos. RNA foi extraído das amostras, e o método de reação em cadeia da polimerase via transcriptase reversa quantitativa (RT-qPCR) foi utilizado para investigar os vírus. A paciente apresentou dois quadros clínicos distintos. O primeiro foi caracterizado por febre, dor no peito e no corpo, prostração e fadiga, que cessou no nono dia. A RT-qPCR foi positiva apenas para o vírus da influenza A (H3N2). Onze dias após o início dos primeiros sintomas, a paciente apresentou dor de garganta, congestão nasal, coriza, prurido nasal, espirros e tosse. Um segundo teste de RT-qPCR foi positivo apenas para SARS-CoV-2. No segundo evento, os sintomas duraram 11 dias. O sequenciamento do SARS-CoV-2 identificou a cepa Ômicron BA.1. Dentre os contatos da paciente, um teve coinfeção por influenza A (H3N2) e SARS-COV-2 (cepa BA.1.15), e os outros dois foram infectados apenas por SARS-CoV-2 (um também pela cepa Ômicron BA.1.15 e o outro pela BA.1.1). Nossos achados reforçam a importância de testes para a detecção de diferentes vírus em casos de suspeita de infecção viral respiratória durante a vigilância epidemiológica de rotina, visto que as manifestações clínicas comuns da COVID-19 imitam as de outros vírus, como o vírus influenza.

Safety and immunogenicity of influenza A(H3N2) component vaccine in juvenile systemic lupus erythematosus

Abstract Introduction Seasonal influenza A (H3N2) virus is an important cause of morbidity and mortality in the last 50 years in population that is greater than the impact of H1N1. Data assessing immunogenicity and safety of this virus component in juvenile systemic lupus erythematosus (JSLE) is lacking in the literature. Objective To evaluate short-term immunogenicity and safety of influenza A/Singapore (H3N2) vaccine in JSLE. Methods 24 consecutive JSLE patients and 29 healthy controls (HC) were vaccinated with influenza A/Singapore/ INFIMH-16-0019/2016(H3N2)-like virus. Influenza A (H3N2) seroprotection (SP), seroconversion (SC), geometric mean titers (GMT), factor increase in GMT (FI-GMT) titers were assessed before and 4 weeks post-vaccination. Disease activity, therapies and adverse events (AE) were also evaluated. Results JSLE patients and controls were comparable in current age [14.5 (10.1-18.3) vs. 14 (9-18.4) years, p = 0.448] and female sex [21 (87.5%) vs. 19 (65.5%), p = 0.108]. Before vaccination, JSLE and HC had comparable SP rates [22 (91.7%) vs. 25 (86.2%), p = 0.678] and GMT titers [102.3 (95% CI 75.0-139.4) vs. 109.6 (95% CI 68.2-176.2), p = 0.231]. At D30, JSLE and HC had similar immune response, since no differences were observed in SP [24 (100%) vs. 28 (96.6%), p = 1.000)], SC [4 (16.7%) vs. 9 (31.0%), p = 0.338), GMT [162.3 (132.9-198.3) vs. 208.1 (150.5-287.8), p = 0.143] and factor increase in GMT [1.6 (1.2-2.1) vs. 1.9 (1.4-2.5), p = 0.574]. SLEDAI-2K scores [2 (0-17) vs. 2 (0-17), p = 0.765] and therapies remained stable throughout the study. Further analysis of possible factors influencing vaccine immune response among JSLE patients demonstrated similar GMT between patients with SLEDAI < 4 compared to SLEDAI ≥ 4 ( p = 0.713), as well as between patients with and without current use of prednisone ( p = 0.420), azathioprine ( p = 1.0), mycophenolate mofetil ( p = 0.185), and methotrexate ( p = 0.095). No serious AE were reported in both groups and most of them were asymptomatic (58.3% vs. 44.8%, p = 0.958). Local and systemic AE were alike in both groups ( p > 0.05). Conclusion This is the first study that identified adequate immune protection against H3N2-influenza strain with additional vaccine-induced increment of immune response and an adequate safety profile in JSLE. ( www.clinicaltrials.gov , NCT03540823).

Efficacy of induction chemotherapy or chemoradiotherapy followed by surgery versus definitive chemoradiation in stage ⅢA-N2 non-small cell lung cancer: A systematic review and meta-analysis / 中国胸心血管外科临床杂志

@#Objective    To systematically evaluate the efficacy of neoadjuvant chemoradiotherapy or chemotherapy followed by surgery versus definitive chemoradiation in stage ⅢA-N2 non-small cell lung cancer (NSCLC). Methods    We searched PubMed, EMbase, Web of Science and The Cochrane Library to collect clinical studies on the efficacy comparison between neoadjuvant chemoradiotherapy or chemotherapy followed by surgery and definitive chemoradiation in stage ⅢA-N2 NSCLC from inception to September 2022. The meta-analysis was performed by using RevMan 5.3 software. Results    A total of 9 studies (3 randomized controlled trials and 6 retrospective cohort studies) with 12 801 patients were included. The results of meta-analysis showed that there was no statistical difference in the progression-free survival rate between the inductive treatment followed by surgery (including lobectomy and pneumonectomy) and definitive chemoradiation (HR=0.99, 95%CI 0.86-1.15, P=0.91). Compared with definitive chemoradiation, the overall survival (OS) rate in the inductive treatment followed by surgery (including lobectomy and pneumonectomy) was lower (HR=1.24, 95%CI 1.09-1.42, P=0.001), while the OS rate in the inductive treatment followed   by lobectomy was higher (HR=0.55, 95%CI 0.51-0.61, P<0.000 01). And the local recurrence rate in the inductive treatment followed by surgery was reduced (OR=0.44, 95%CI 0.36-0.55, P<0.000 01). Conclusion    Neoadjuvant chemoradiotherapy or chemotherapy followed by lobectomy is superior to definitive chemoradiation in OS and it has a lower local recurrence rate, so lobectomy should be one of the multidisciplinary treatments for selected ⅢA-N2 NSCLC patients.

Sequence analysis of HA and NA genes of human infected H9N2 avian influenza virus in Yunnan province, 2019 / 中国热带医学

@#Abstract: Objective To investigate the molecular characteristics of the H9N2 avian influenza virus (AIV) causing human infection in Yunnan Province in 2019, and to provide the scientific basis for the prevention and control of avian influenza in Yunnan Province. Methods Influenza virus typing was performed by real-time RT-PCR in two influenza-like illness samples, and the Illumina Miseq high-throughput sequencer was used to determine the viral genome sequence. HA and NA gene sequence alignment and phylogenetic tree construction were performed using Mega7.0 software. Results Real-time RT-PCR results showed that two influenza-like illness samples were positive for H9N2 subtype. The full length of HA and NA were obtained by genomic sequencing. Sequence system evolution analysis showed that the HA and NA of the two AIVs in Yunnan Province were in the same evolutionary clade as A/Chicken/Zhejiang/HJ/2007 and belonged to the G57 type. The HA nucleotide and amino acid homology of the two AIVs were 93.92% and 95.00%, respectively, and the NA nucleotide and amino acid homology was 93.31% and 82.03%, respectively. The nucleotide (amino acid) homology of HA was 92.29%-96.94% (93.77%-98.43%) and 92.84%-94.92% (94.18%-96.23%), respectively, and NA nucleotide homology (amino acid) were 91.81%-97.60% (77.82%-94.83%), 94.38%-97.22% (85.47%-94.55%), respectively, compared with that of human infected H9N2 epidemic strains obtained in China from 2015 to 2020. Both AIVs HA protein cleavage site sequences were PSRSSR↓GLF, which was in line with the characteristics of low pathogenic influenza. The analysis of HA protein receptor binding site showed that amino acids at positions 109, 161, 163, 191, 202, 203 and 234 were consistent with the reference strains, while amino acids at position 198 were mutated to T. N166D and 168N mutations were also found in HA protein, and both AIVs had 7 potential glycosylation sites. Analysis of the erythrocyte binding site of NA gene found that there were amino acid mutations at positions 369, 402, 403, and 432, and amino acid deletion at positions 63-65 was found in the NA genes. There were 4 and 5 potential glycosylation sites in the two AIVs, respectively, and no drug resistance site mutations were found. Conclusions　The receptor binding sites, erythrocyte binding sites and glycosylation sites of HA and NA genes of H9N2 AIV in Yunnan Province have different degrees of variation, and monitoring and prevention and control should be strengthened.

Research progress on efficacy and influencing factors of postoperative radiotherapy for stage ⅢA(N 2) non-small cell lung cancer / 中华放射肿瘤学杂志

Non-small cell lung cancer(NSCLC) is highly malignant and has poor prognosis, in which stage ⅢA(N 2) NSCLC approximately accounts for 20%. Patients with stage ⅢA(N 2) NSCLC have high heterogeneity and distinct survival difference. Loco-regional recurrence and distant metastasis are the main causes of treatment failure. At present, whether stage ⅢA(N 2) NSCLC patients should receive postoperative radiotherapy(PORT) remains controversial. Such patients still lack high level proof to receive PORT.

Expression and significance of fork head box protein N2 and F2 in gliomas / 解剖学报

Objective To observe the expression levels of fork head box protein N2 (FOXN2) and fork head box protein F2 (FOXF2) in normal brain tissue, low-grade glioma and high-grade glioma, and to explore the relationship between the two and the grade of glioma, and then deduce their roles in the occurrence and development of glioma, to look for possibilities target of drug therapy. Methods From January 2016 to December 2020, 36 glioma specimens were collected from the Department of Pathology, Yijishan Hospital, Wannan Medical College. The expressions of F0XN2 and F0XF2 were detected by immunohistochemistry and immunofluorescence with the double-blind method, and analyzed by SPSS18. 0 software, with a statistical significance of P<0. 05. Then Western blotting and Real-time PCR experiments were carried out on fresh normal brain tissues, low-grade glioma tissues, high-grade glioma tissues during neurosurgery in Yijishan Hospital of Wannan Medical College from January 2020 to January 2021 (normal brain tissues were all trauma patients). Results The results of immunohistochemistry and immunofluorescence showed that the higher grade of glioma, the lower expressions of FOXN2 and FOXF2 would be (P<0. 05). Western blotting showed that the expressions of FOXN2 and FOXF2 in normal brain tissue and low-grade glioma were higher than those in high-grade glioma (P<0. 05). The result of Real-time PCR showed that the expressions of FOXN2 and FOXF2 in normal brain tissue were higher than those in high-grade glioma (P<0. 01). Conclusion FOXN2 and FOXF2 are expressed in normal brain tissues, and their expression is low in glioma in a grade-dependent manner, suggesting that FOXN2 and FOXF2 are related to the grade and poor prognosis of glioma. Enhancing the expression of F0XN2 and F0XF2 ma)' provide a new idea for target therapy of glioma.

Spatio-Temporal history of H9N2 viruses in Iran and neighbor countries by Bayesian analysis and molecular characterization

Objective: To delineate the H9N2 influenza virus circulation within Iran and its neighboring countries, the potential source of the epidemic in these countries, and its date of origin. Methods: We obtained all hemagglutinin (HA) and neuraminidase (NA) nucleotide sequences of influenza H9N2 available up to December 25, 2020 from Iran and its neighboring countries (i.e., Pakistan, Afghanistan, Turkmenistan, Armenia, Azerbaijan, Turkey, and Iraq). We also performed a Bayesian Markov chain Monte Carlo method to infer the evolutionary dynamic and the most recent common ancestor for the HA and NA sequences. Results: H9N2 epidemic may have started in Iran and Pakistan much earlier than the other investigated countries in the region, and an ongoing bidirectional dispersion of the virus between the investigated countries was also observed. The mean time of the most recent common ancestor of H9N2 viruses was 1988 for HA, and 1992 for NA. Conclusions: Strains from investigated countries rooted in Pakistan and Iran. Regular surveillance of H9N2 viruses, especially in the live bird markets, enhancing the biosecurity of poultry industry and screening newly arriving immigrants and tourists from neighboring countries at border should be considered to control spread of the virus. Furthermore, surveillance of viral molecular evolution should be initiated for effective prevention of epidemic and pandemic spreads.

Analysis of hemagglutinin gene characteristics of influenza A H3N2 virus in Changzhou, 2017 -2018 / 公共卫生与预防医学

Objective To understand the epidemiological characteristics and variation of H3N2 influenza virus hemagglutinin (HA) gene in Changzhou from 2017 to 2018. Methods Throat swab specimens of the influenza-like cases were collected from Changzhou Influenza Monitoring Sentinel Hospital from April 2017 to March 2018. RNA was extracted from the specimens for influenza diagnosing and genotyping using real-time RT-PCR．H3N2 positive samples were isolated, and extracted RNA was used for amplification, sequencing and phylogenetic analysis of HA gene. Results From April 2017 to March 2018, 28 strains of influenza A (H3N2) virus were isolated. After gene sequencing, a phylogenetic tree was constructed. It was found that all of the strains belonged to Group3C.2a, which was similar to the vaccine strain A/Hong Kong/4801/2014. The HA amino acid sequence difference was analyzed and compared between the H3N2 influenza virus strains isolated in Changzhou and the vaccine strain A/Hong Kong/4801/2014. It was found that the epidemic strain isolated in Changzhou was in the HA epitope (A-E) region. Ten amino acid site mutations in the HA epitope (A-E) region and two amino acid site mutations in the stem region of HA antigen were found. Conclusion From April 2017 to March 2018, the influenza virus H3N2 prevalent in Changzhou was distributed on the same evolutionary branch with the vaccine strain A/Hong Kong/4801/2014 (group 3C.2a), rendering the popular trend of one subgroup. However, some amino acid sites of the HA epitope had variations, suggesting that mutations may occur, which may affect the immune effect of the vaccine. Monitoring needs to be strengthened in the future work.

Genetic characteristics of hemagglutinin gene of H9N2 avian influenza viruses isolated from live poultry markets in Anhui Province from 2013 to 2018 / 中华疾病控制杂志

@#Objective To analyze genetic characteristics of hemagglutinin( HA) gene of H9N2 avian influenza viruses( AIVs) circulating in Anhui Province from 2013 to 2018． Methods All H9N2 positive samples tested by real－time polymerase chain reaction( PCＲ) were inoculated into specific patho- gen free ( SPF) chicken eggs for isolation and purification． Viral ＲNA was reversely transcribed into cD- NA and then amplified with gene specific primers． PCＲ products were sequenced and the gene sequences were analyzed using molecular and bioinformatics software． The DATAMONKEY online server was conducted to analyze selection pressure，and protein structure homology modelling was computed by the SWISS－MODEL server． Ｒesults 33 H9N2 AIVs isolated from live poultry markets belonged to h9．4．2．5 in the phylogenetic tree． The receptor binding sites of HA gene at 183，226 and 227 position were mutated into N，L and M，respectively． Meanwhile 189 and 190 sites presented with genetic polymorphism． Since 2015，all H9N2 viruses in this study carried 6 potential N－linked glycosylation sites． It was found that po- sition 160 of HA gene was subjected greater positive selection pressure，presented 7 spatial conformations at least． Conclusions The H9N2 viruses isolated from live poultry markets in Anhui Province possess the molecular characteristics of infecting mammals and the ability of antigenic drift，so we need to pay more attention to the genetic characteristics of the viruses．

Respuesta de Fragaria mexicana y comunidades microbianas rizosféricas al aumento de temperature / Response of Fragaria Mexicana and rhizophere microbial communitites to temperature increase

Resumen Para los próximos 100 años se pronostica un incremento en la temperatura del planeta de casi 4 °C, lo cual pondrá en riesgo las especies que no logren adaptarse. En esta investigación se determinaron las respuestas morfofisiológicas de F. mexicana y los cambios en la población de bacterias fijadoras de nitrógeno atmosférico (BFN) asociadas a sus raíces, debido a dos condiciones ambientales con diferencias medias de temperatura de 5.1 oC: invernadero (temperatura alta, TA) y campo abierto (temperatura baja, TB); y con cuatro tratamientos: sin inocular (T), inoculadas con la cepa ocho (CP8), cepa cuatro (CP4) y con ambas cepas (CP8 + 4). Las BFN fueron aisladas de la rizosfera de F. mexicana y transformadas genéticamente con reporteros, para cuantificar la población al final del experimento. Se midió el peso seco de la parte aérea y la raíz, la tasa de asimilación de CO2 (A), el inicio de la floración, el número de flores y frutos; y la persistencia de las bacterias fijadoras de N atmosférico (BFN). Además, se evaluó la concentración de NO3, NH4, P y materia orgánica (MO) en el sustrato, al inicio y final del experimento. Las plantas sometidas a alta temperatura presentaron mayor peso seco de la parte aérea y fotosíntesis; con una disminución en el peso seco de la raíz, y en el número de flores y frutos. La MO en el sustrato disminuyó, mientras que la disponibilidad de NO3, NH4 y P aumentó. El incremento de temperatura y la mayor presencia de N en el substrato provocaron reducción en la población de BFN. Estos resultados sugieren que temperaturas altas estimulan el crecimiento de F. mexicana y tienen un impacto negativo sobre su reproducción y en las BFN asociadas a sus raíces.(AU)

Abstract The earth could experience a warming of 4 °C in the next one hundred years. This would put at risk the plants that can´t adapt. Fragaria mexicana is an endemic plant of temperate forest of Mexico. The response of this wild strawberry to temperature increasing has not been studied and could play an important role for event of global warming. This study determinate the morphological and physiological responses of F. mexicana and changes in the N2-fixing bacteria (BFN) population on its roots, due to two environmental conditions with differences of temperature 5.1 °C: greenhouse (high temperature, TA) and open land (low temperature, TB); and with four free-living nitrogen-fixing bacteria (BFN) treatments: non-inoculated (T), inoculated with strain eight (CP8), strain four (CP4), and strains eight and four mixed (CP8 + 4). BFN were isolated from the rhizosphere of F. mexicana and were genetically transformed with reporters to quantify the population at the end of the experiment. NO3, NH4 and P and organic matter (MO) in the substrate were determined at beginning and finish of the experiment. Shoot and root dry weight, photosynthetic rate, flowering and fructification starting, flowers and fruits number, were measured. Shoot dry weight and photosynthetic rate were lower in TB than TA, decreasing 3.1 g and 0.94 µmol m-2 s-1, respectively. Root dry weight was 3.0 g less in TA compared with TB. Number flowers decrease in 40.89 % and number fruits in 38.11 % on TA than TB. F. mexicana plants start flowering 14 days previously in TB than TA. MO in the substrate decrease in TA, while the concentration of NO3, NH4 and P, increased. Population of BFN was lower in TA. Results obtained indicated that higher temperature promotes the growth of F. mexicana and reduce its reproduction and BNF population associated with its roots.(AU)

Molecular evolution of influenza A/H3N2 viruses HA and NA genes circulating in Shenzhen in 2017 / 中华疾病控制杂志

Objective To investigate the molecular evolution of the Hemagglutinin (HA) and Neuraminidase (NA) genes of influenza A/H3N2 viruses in Shenzhen in the first half of 2017, so as to provide scientific basis for predicating influenza epidemic and variation. Methods A total of 40 influenza A/H3N2 viruses strains were selected and the molecular phylogenetic trees were constructed by bioinformatics software DNAStar, MEGA 7.0, etc. Then, the genetic characteristics and variation of HA and NA genes along with corresponding amino acids were analyzed. Results The homology of Shenzhen isolates reached 97.8%-100.0%, which located in the human-derived branch of Asia and North America. Compared with the vaccine strains A/Switzerland/9715293/2013(H3N2) and A/Hong Kong 14801/2014(H3N2) recommended by world Health Oraganication (WHO), there was a higher sequence similarity. Compared with the vaccine strain, HA and NA proteins had a number of amino acid sites replaced, of which HA 6 antigen sites and 2 receptor binding sites change; NA had a mutation of D151N/G located in enzyme activity sites. Potential N-glycosylation sites for HA and NA also changed. Conclusions The influenza A/H3N2 viruses in Shenzhen in the first half of 2017 has not yet formed a new subtype in the epidemic. Currently, the recommended vaccine strains still have some protective effects on the population. The replacement mutation of multiple amino acids sites of HA and NA suggests that the dynamic monitoring of molecular level of influenza A/H3N2 viruses need to be strengthened.

Study on the source of human infection with H9N2 influenza virus from live poultry markets in Changsha / 中华疾病控制杂志

Objective To explore the source of human infection H9N2 avian influenza virus (AIV). Methods Environmental AIV nucleic acid monitoring was conducted for live poultry markets in Changsha city from 2014 to 2015, and data of human infection H9N2 subtype AIV cases worldwide were collected. Phylogenetic trees of hemagglutinin(HA), neuraminidase(NA)and non-structural protein(NS)genes from human infection H9N2 subtype AIV, the live poultry markets environmental H9N2 subtype AIV and partial avian H9N2 subtype AIV were constructed using the MEGA 6.06 software, respectively. Results In 2014-2015, H9 subtype AIV had the highest nucleic acid positive rate (44.76%) in the live poultry markets environment of Changsha city, and the pollution was serious. A total of 27 cases of human infection with H9N2 subtype AIV had been reported worldwide, and most of these patients recovered after treatments.Epidemiological survey showed that 59.26% (16/27) of cases had a clear history of exposure to poultry or live poultry markets. The phylogenetic trees of HA, NA and NS genes showed that the human infection H9N2 subtype AIV isolates isolated from Hunan and Guangdong were closely related to the H9N2 subtype AIV isolated from the live poultry markets environment in Hunan and Guangdong provinces from 2013 to 2016. The nucleotide similarity was as high as 97%-99%. Conclusion Live poultry market is one of the sources of human infection with H9N2 influenza virus.

Inhibition of the replication of H9N2 influenza virus in vivo by short-term repeated oral administration of chicken interferon α / 生物工程学报

To evaluate the optimal administration frequency for interferon-α (IFN-α) and the effect of its combined use with inactive virus on chicken flocks, the prokaryotic expression plasmid pET-22b-ChIFN-α was constructed and transferred into Escherichia coli BL21(DE3) host bacteria to induce the expression of chicken IFN-α and to harvest recombinant proteins inclusion bodies. The expression of recombinant chicken IFN-α was confirmed by SDS-PAGE, and the results demonstrated that the chicken IFN-α (20 kDa) was highly expressed using the prokaryotic expression vector with a concentration of 0.2 mg/mL in the medium. Chicken IFN-α was diluted to 2.5×10⁴ U/fowls and administered to immunized specific-pathogen-free chickens orally in combination with inactivated H9N2 subtype influenza virus. Chicken that received chicken IFN-α were safe after three repeated immunizations (96 h). In addition, chicken IFN-α could induce higher levels of antiviral-related inducible genes in peripheral blood, spleen, and thymus of chicken flocks. The results of a challenge assay revealed that the lowest detoxification rates of chicken IFN-α ranged from three to five days, suggesting a higher capacity to resist H9N2 subtype avian influenza virus. The present study obtained the optimal immune frequency and immunization period for chicken IFN-α to provide theoretical support for the optimal clinical application of IFN-α.