Objective:To study and establish the UPLC fingerprint and multi-index content determination methods of Hedyotidis chrysotricahae Herba; To provide a reference for the quality control of Hedyotidis chrysotricahae Herba.Methods:The chromatographic column was ACQUITY UPLC HSS T3 (100 mm×2.1 mm,1.8 μm). The mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution; the detection wavelength was 254 nm; the flow rate was 0.30 ml/min and column temperature was 35 ℃. The method could determine content and fingerprint of rutin, Kaempferol-3-O-rutinoside, Narcissoside, Neochlorogenic aci, Chlorogenic Acid, Cryptochlorogenic acid and have quality analysis to 17 batches of Hedyotidis chrysotricahae Herba based on the variance of fingerprint, similarity evaluation, clustering analysis along with principal component analysis (PCA) at the same time.Results:The common pattern of UPLC specific chromatogram of Hedyotidis chrysotricahae Herba was established. The 11 common peaks were marked out, among which 7 peaks were identified. 17 batches Hedyotidis chrysotricahae Herba could be divided into 4 categories according to different origins. Quality content of six indicators of Hedyotidis chrysotricahae Herba was in slight difference between different origins, among which the content quality of Hedyotidis chrysotricahae Herba from Duyun in Guizhou Province was the highest.Conclusion:The established UPLC fingerprint and content determination method of 6 indicators from the study can be used for the quality control of Hedyotidis chrysotricahae Herba, which can also provide a theoretical basis for the standard improvement of Hedyotidis chrysotricahae Herba.

Objective To conduct a unique and pioneering molecular epidemiological investigation of a case of Eurasian avian-like H1N1 swine influenza identified in Yunnan Province in 2022(the fourth such case in the province)and to understand its genetic characteristics so as to reveal its potential impact on human health.Methods Real-time fluorescent quantitative PCR detection technology was used for the nucleic acid testing of the case's pharyngeal swab samples,close contacts,and environmental samples from the living area.Positive samples were subjected to virus isolation using MDCK cells.Cell cultures were authenticated using erythrocyte agglutination assay with guinea pig blood and real-time fluorescence quantitative RT-PCR.Whole genome sequencing was performed using the Illumina MiseqNext-generation sequencing platform,and a phylogenetic tree was constructed using MEGA 7.0 software to analyze the genetic molecular characteristics.Results The first G5 genotype Eurasian avian-like H1N1 swine influenza virus in Yunnan Province was successfully isolated,and the whole genome sequence of the virus was obtained.This virus possessed the molecular characteristics associated with increased adaptability,virulence,or transmissibility in mammals and had a nucleotide consistency of 99.2％～99.7％with a porcine strain isolated in Jiangsu province.These findings underscored the potential threat this virus poses to human health.Conclusion The study underscores the importance of further monitoring swine influenza in preventing new influenza virus subtypes that can infect humans.

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.

Objective:To understand the characteristics of the local Coronavirus Disease 2019 (COVID-19) epidemic in the border areas of Yunnan province, and to analyze and trace the genome variation of the severe acute respiratory syndrome coronavirus 2 (SARS-COV-2).Methods:The basic information and samples of the first cases of 12 local epidemics in the border areas of Yunnan province from mid-February to April 2022 were collected. The virus genomes, genotypes and mutations were determined by the whole genome sequencing technology, the Pangolin COVID-19 Lineage Assigner and MEGA software. The sources of virus infection were estimated based on the contents of epidemiological investigations.Results:The first cases from 12 local epidemics in the border areas of Yunnan province had diverse occupations, and none of them had ever left the local area. The virus genomes of 12 first cases were Omicron (BA.2) variants and every two of them further belonged to the evolutionary clades BA.2.10 and BA.2.3.2, respectively. The genome-wide nucleotide-level mutation analysis showed that there were 1 or more specific nucleotide mutation sites in 10 first cases of the local epidemics. Combined with the epidemiological investigations, it was suggested that there might be different sources of virus infection in all 12 local epidemics. Further analysis of amino acid mutations revealed that in addition to the representative mutation sites of evolutionary clades, there were also specific amino acid mutation sites (such as S: E1188 D) in the genome sequences between the cases, which also deserved attention. The result of phylogenetic tree analysis were consistent with Pangolin Assigner and mutation analysis.Conclusions:The 12 local epidemics in Yunnan may have different sources of virus infection, which indicates that the COVID-19 epidemic in the border areas of Yunnan province is extremely complicated. Strengthening the frequency of nucleic acid screening and the scope of key populations and monitoring the variation of SARS-CoV-2 are of great significance for the precise prevention and control of the epidemic.

Objective:To understand the viral spectrum changes of respiratory tract infections in Kunming city from 2018 to 2021.Methods:Throat swab samples were collected from influenza-like cases in 2 general hospitals and 1 children’s hospital in Kunming city from 2018 to 2021. Real-time RT-PCR was used to detect 14 common respiratory viruses (including influenza virus, human adenovirus, human respiratory syncytial virus, human rhinovirus, human bocavirus, human metapneumovirus, human parainfluenza virus, human coronavirus), and the detection data before and during the epidemic were analyzed.Results:From 2018 to 2021, a total of 12 302 specimens were collected and tested, and 2 600 positive specimens were detected, with a total positive rate of 21.13%. The positive detection rate before the epidemic was 25.91%, which was significantly higher than 16.18% during the epidemic ( χ2=174.35, P<0.05), and the detection rates of Flu and HAdV decreased by 75.36% and 45.31% respectively, while the detection rates of HRV and 229E increased by 72.22% and 350% respectively. During the epidemic, the total virus detection rate of male and female cases and all age groups under 50 years old were lower than before the epidemic. Conclusions:The epidemic of SARS-CoV-2 and the implementation of corresponding prevention and control measures have an impact on the epidemic trend of common respiratory viruses in Kunming, and the monitoring of multiple respiratory viruses is of great significance.

Objective:By analyzing the viral pathogen spectrum and epidemiological characteristics of hospitalized severe acute respiratory infection (SARI) cases in Kunming city to provide the pathogenic basis for the clinical diagnosis, treatment, prevention and control of acute respiratory tract infection.Methods:Respiratory viruses were detected by real-time PCR, and the results were statistically analyzed using software SPSS20.0.Results:A total of 291 SARI cases’ pharyngeal swab specimens were detected from January 2018 to March 2019, and 86 positive specimens were detected, the positive rate was 29.55%. The top three pathogens detected were human influenza A virus (11.34%), human metapneumovirus (3.44%), and human respiratory syncytial virus (3.09%), and there were 6 specimens with mixed infection of two viruses, and the mixed infection rate was 2.06%. The virus detection rate of male cases was higher than that of female cases ( χ2=6.183, P=0.013), and the positive rate of virus detection was the highest in the 2 to 5 years age group, and positive respiratory virus detection peaks were in the fourth and the first quarter. Comparison of virus positive and negative group showed that there were no statistically significant differences in clinical symptoms, signs, complications and underlying diseases between the two groups except for expectorant cough ( χ2=5.107, P=0.024) and runny nose ( χ2=4.683, P=0.030). Conclusions:Influenza virus is the main pathogen of SARI cases in Kunming city, and the detected pathogens have certain epidemic rules in different age groups and seasons.

Objective: To explore the current situation of nurses′ working values, working environment and head nurses′ leadership style. To explore the influence factors of nurses′ working environment and head nurse′s leadership style on nurses′ working values. Methods: By applying random stratified sampling, 499 clinical nurses without administrative titles in 6 hospitals were selected. Questionnaires were adopted as the main research tool. Results: Score of nurses′ working values was 3.52 ± 0.56. Score of nurses′ working environment was 3.03 ± 0.44. Score of head nurses′transformational leadership style was 2.70 ± 0.76, and score of head nurses′ transactional leadership style was 2.23 ± 0.47. Working environment, transformational leadership style and transactional leadership style were positively correlated with nurses′ working values (P= 0.452, 0.371, 0.234). Conclusions Working values of nurses in general public hospitals of Changsha city is at moderate level. By improving nurses′ working environment and head nurses adopting positive leadership, and in particular, proposing beneficial measures for improving a solid nursing foundation for quality of care, sufficient human resources and intellectual stimulation, nurses′ working values can be improved.

Aortic dissection is a life-threatening cardiovascular condition. The elevated blood pressure plays an important role in the development and the formation of aortic dissection, thus treatment of aortic dissection requires the management of blood pressure control. In this paper, we reported the current situation and summarized the influencing factors of blood pressure management in the treatment of patients with aortic dissection. Suggestions were provided to improve the management of blood pressure control and to support the future research in China.

Objective To express and purify the recombinant UL7 protein of herpes simplex virus 1 (HSV-1), to prepare the corresponding UL7-specific polyclonal antibody and to preliminarily analyze the expression of UL7 protein during the proliferation of HSV-1. Methods The UL7 gene was amplified by PCR and then cloned into the pGEX-5X-1 vector for expression of UL7 protein in the prokaryotic expression system. The constructed expression plasmid, pGEX-5X-1-UL7, was transformed into E. coli BL21 (DE3) to induce the expression of UL7 protein by IPTG. The purified GST-UL7 fusion protein was used as antigen to inject the ICR mouse for the preparation of polyclonal antibody specific for UL7 protein. The titer and speci-ficity of the polyclonal antibody were analyzed by using indirect ELISA and Western blot assay, respectively. The UL7 protein-specific polyclonal antibody was used to detect the expression of UL7 protein at different time points after infecting Vero cells with HSV-1. Results The GST-UL7 fusion protein was efficiently ex-pressed in E. coli BL21 (DE3). The UL7 protein-specific polyclonal antibody was prepared with high titer (1 ∶ 105) and high specificity as indicated by the indirect ELISA and Western blot assay. The expression of UL7 protein was detected at different time points after infecting Vero cells with HSV-1. Conclusion The GST-UL7 fusion protein was obtained successfully and the UL7 protein-specific polyclonal antibody was pre-pared. Accompany with the proliferation of HSV-1, the expression of UL7 protein was detected at different time points by using the polyclonal antibody.