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.

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 investigate the prevalence and influencing factors of soil-transmitted nematode infections in Ningbo City from 2016 to 2021, so as to provide insights into the development of targeted control measures against soil-transmitted nematodiasis. Methods Permanent residents at ages of 3 years and older were sampled in Ningbo City using a multi-stage sampling method each year during the period from 2016 to 2021. Soil-transmitted nematode eggs were detected in stool samples using a modified Kato-Katz thick smear method (two slides for each stool sample), and Enterobius vermicularis eggs were additionally identified among children at ages of 3 to 12 years using the adhesive cellophane-tape perianal swab method. The time- and regions-specific prevalence of soil-transmitted nematode infections was calculated, and the factors affecting hookworm infections were identified using a multivariate logistic regression model. Results A total of 11 573 person-times were detected for soil-transmitted nematode infections in Ningbo City from 2016 to 2021, and 296 egg-positives were detected, with a mean prevalence rate of 2.56% [95% confidential interval (CI): (2.28%, 2.87%)]. Hookworm was the predominant species of soil-transmitted nematode among egg-positives in Ningbo City (98.31%, 291/296), and there was a significant difference in the prevalence of hook-worm infections among years (χ² = 190.27, P < 0.01). The highest prevalence of hook-worm infections was observed in Ninghai County (4.06%), and there was a region-specific prevalence rate of hookworm infection in Ningbo City (χ² = 148.43, P < 0.01). Multivariate logistic regression analysis showed that elderly residents at ages of over 60 years [odds ratio (OR)= 1.94, 95% CI: (1.07, 3.54), P < 0.05], males [OR = 2.19, 95% CI: (1.72, 2.80), P < 0.01], farmers [OR = 6.94, 95% CI: (3.37, 14.29), P < 0.01] and residents with a low education level [illiteracy or semi-illiterate: OR = 3.82, 95% CI: (1.56, 9.35), P < 0.05; primary school: OR = 2.70, 95% CI: (1.11, 6.59), P < 0.05] were at a higher risk for hookworm infections. Conclusions The overall prevalence of soil-transmitted nematode infections was low among residents in Ningbo City from 2016 to 2021. The surveillance and health education for human hookworm disease remain to be reinforced among male farmers with a low education level at ages of over 60 years in Ninghai County.

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: 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.

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.

Objective To investigate the prevalence of thyroid nodules among residents of Ningbo City and to explore the risk factors for thyroid nodules.Methods Totally 3 596 residents aged 6-70 were selected by multistage stratified cluster random sampling in Ningbo City,2011.Thyroid ultrasound examination was performed in all participants,while salt iodine and urinary iodine were measured.Possible risk factors for thyroid nodules were analyzed by single factor and multi-factor logistic regression.Results The prevalence rate of thyroid nodules in residents of Ningbo City was 22.66％ (815/3 596),with standardized prevalence rate of 17.69％.Female [(odds ratio (OR) =2.06,95％ confidence interval (CI) =1.74-2.43],groups of aged 18-(OR =3.42,95％ CI =1.22-9.61),aged 40-(OR =13.06,95％ CI =4.80-35.51),aged 65-(OR =16.67,95％ CI =5.97-46.54) and occasional consumption of seafood habits (less than twice per week,OR =1.23,95％ CI =1.02-1.49) were significant associated with thyroid nodules.Conclusions There is no link Letween iodine nutrition and thyroid nodules.It is demonstrated that female and aged ≥ 18 groups have higher risk of thyroid nodules; frequent consumption of seafood seems to be a protective factor.

Objective To investigate the iodine nutritional status,thyroid function and the prevalence of thyroid nodules of residents using non-iodized salt in Ningbo City,Zhejiang Province.Methods In 2011,using typical sampling method,6 villages/communities consumption of non-iodized salt were selected as study fields and residents aged 6-70 were selected as study subjects in Ningbo.Urinary iodine detection,thyroid ultrasound examination and tests of thyroid function [free triiodothyronine (FT3),free thyroxine (FT4),thyroid stimulating hormone (TSH),thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb)] were performed in all participants.Urinary iodine concentration (UIC) was determined by arsenic cerium catalytic spectrophotometry,and thyroid function was monitored by chemiluminescence method.Results A total of 1 177 residents agreed to participate in the study,482 were male,and 695 were female (including 2 pregnant women).The participants' mean age was (48.54 ± 13.98) years.The median UIC in all participants was 90.36 μg/L,and the proportions with a UIC ＜ 100μg/L was 56.07％ (660/1 177).Furthermore,the median UIC in male was 97.05 μg/L and in female was 83.84 μg/L,there was a significant difference in UICs between male and female (Z =-2.99,P ＜ 0.05).Similarly,significant difference in UICs was observed among age groups (x2 =99.34,P ＜ 0.01).Among the participants,19.88％ had thyroid nodules,with standardized prevalence rate of 13.95％.Furthermore,in male,11.83％ had thyroid nodules,with standardized prevalence rate of 8.11％,and in female was 25.47％,with standardized prevalence rate of 19.83％,there was a significant difference in the detection rate of thyroid nodules between male and female (x2 =33.25,P ＜ 0.01).Likewise,significant difference in the detection rate of thyroid nodules was observed among age groups (x2=49.65,P＜ 0.01),and there was a positive and significant correlation between the detection rate of thyroid nodules and increasing age (x2trend =44.22,P ＜ 0.01).Conclusions The residents who using non-iodized salt in Ningbo City are in the status of iodine deficiency.The detection rate of thyroid nodules is higher in female than in male and it has increased with age.