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 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.

Traditional Chinese Medicine (TCM) therapy is effective in treating malignant tumor. TCM therapy is a multi-way, multi-layer and multi-target integrated treatment characterized by "survival with tumor" and improvement of life quality. An ideal result of the TCM therapy for cancer should be a positive response of the patient with satisfactory quality of life (QOL) and longer survival time. So it is necessary to introduce the QOL, quantification of symptoms and signs and standardization of TCM syndromes to the objective, quantitative and standardized effect assessment of TCM therapy for cancer. A criterion for evaluating the effect of TCM therapy on tumors should be established, which reflects the characteristics and advantages of the TCM therapy, and meets the requirement of the developing effect assessment of modern medicine in cancer treatment. It is suggested that the treatment effect should be the sum of quality adjusted life years (QALYs) (equal to QOL multiplied by time of survival), syndrome remission rate and tumor remission rate. But the proportion of each of the three is variable, which has close relationships with the different tumor stages and corresponding treatments.

Objective: To understand the concentrations and source of SO42-,NO3-,Cl- and NH4+ in atmospheric PM2.5 in Zhoushan,and to provide reference for controlling PM2.5 and formulating effective environmental protection measures. Methods: Monitoring sites in new districts of Zhoushan were set up to continuously collect PM2.5 from 10th to 16th of each month and under the hazy weather during 2015-2016（AQI > 200）. The mass concentration of PM2.5 was measured by weighing method,and the concentrations of SO42-,NO3-,Cl- and NH4+ in PM2.5 components was detected by ion chromatography. Results: The average daily concentration of PM2.5 in Zhoushan from 2015 to 2016 was（40.91±27.39）μg/m3. The concentration of the four water-soluble non-metal ions in PM2.5 components was 3.56-103.03 μg/m3,with an average of（23.06±20.00）μg/m3,accounting for about 56.64% of PM2.5 contents. The average monthly concentration of SO42- was the highest[（10.35±6.48）μg/m3],while the average monthly concentration of Cl- was the lowest [（0.49±0.73）μg/m3]. The concentration of the four ions was the highest in winter[（37.56±27.74）μg/m3]and the lowest in summer[（12.32±5.88）μg/m3]. The differences between different seasons was statistically significant（P<0.05）. The highest concentration of NO3- occurred in winter,which was（14.48±13.28）μg/m3. The concentration ratio of NO3- to SO42- ranged from 0 to 2.58,with an average of 0.55. There were 28 days（14.74%）with the ratio greater than one,22 days of which was in winter. Conclusion The concentration of SO42- was the highest and Cl- was the lowest in atmospheric PM2.5 in Zhoushan. The highest concentration of the four ions occurred in winter. The concentration of NO3- in winter was higher than that of SO42-,suggesting that motor vehicle exhaust might be the main source of PM2.5 in winter.

Chronic ulcer of the lower extremities amounts for a grave and serious problem for public health. Western medicine focuses on controlling infection, improving blood circulation, surgical debridement, skin grafting, etc, but there are bottlenecks in the treatment. Traditional Chinese medicine (TCM) has a long history and a legacy of sound clinical efficacy in this area. TCM has developed a unique, effective external theory, and a large number of topical prescriptions and external technology. Through this research, a safe and effective treatment protocol of TCM for chronic ulcer of the lower extremities can be formed. To this end, during China's "Eleventh Five-Year" Plan, special research committees and projects on TCM external treatments and external technologies were established. This study on ulcer of the lower extremities constitutes one of the major research topics.

OBJECTIVETo study the inhibition effect of winter seedling raising in sunlight-greenhouse on premature bolting of Angelica sinensis.

METHODOne factor of sowing date with 4 levels was tested at random design with 3 repeats.

RESULTSowing date of winter seedling raising had an important effect on seedling growth, seedling mature, plant growth, premature bolting, yield, quality, and extreme significant effect on yield. The bolting date of winter raised seedlings started in middle of July, 50 d later than tranditional seedlings, and bolting peck date was in the last ten days of July, 40 d later than the traditional seedlings. The highest ratio of premature bolting for winter raised seedlings was 1%, and the lowest was 0. A. sinensis roots sowed in November 28 had the highest yield, root dry ratio, ethanol extract, essential oil and ferulic acid contents compared to that in other sowing dates. The best sowing time was from end of November to middle of December.

CONCLUSIONPremature bolting of A. sinensis could be greatly inhibited by winter seedling raising, end of November to middle of December would be the best sowing time for winter seedling raising in greenhouse.

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.

Professor Li Candong puts forward the"five differentiation"thinking in TCM:symptom differentiation,syndrome differentiation,disease differentiation,person differentiation and mechanism differentiation.This article discussed the thinking and method of application of classical prescriptions based on the mode of"five differentiation".Treatment based on symptom differentiation is a quick method of application of classic prescriptions,which includes searching for specific symptoms or symptom groups and according to special tongue images.Treatment based on syndrome differentiation is a commonly used method in classic prescriptions,distinguishing between primary and secondary syndromes and the authenticity of cold and heat.Treatment based on disease differentiation is the inherent meaning of classic prescriptions,which is mainly to distinguish six meridian diseases and special prescriptions for specific diseases.Treatment based on person differentiation embodies the individual differences in the use of classic prescriptions,which include age,gender,constitution and abdominal syndrome.Treatment based on mechanism differentiation is an ingenious method used by classic prescriptions.When practicing clinical medicine,we should adhere to the integrated mode of"five differentiation"in the application of classic prescriptions,comprehensively considering the five dimensions,in order to improve the accuracy and effectiveness of the application of classic prescriptions,reveal and improve the academic system of classic prescriptions,and better guide their clinical application.

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.