AIM:To predict the active compo-nents and targets of Piperlongum L.and the associ-ated signaling pathways involved in pulmonary fi-brosis using network pharmacology and molecular docking technique and evaluate the mechanism of Piperlongum L against pulmonary fibrosis by in vi-tro experiments.METHODS:The active ingredients and targets were retrieved from TCMSP,Swiss Tar-get Prediction and PubChem databases.The dis-ease-related targets were retrieved from Gene-Cards and OMIM databases.The intersection tar-gets of the drugs and disease-related targets were identified using jvenn online tool.String database was used to construct the"drug-component-tar-get"and PPI network and the networks were visual-ized using Cytoscape 3.9.1 software.GO and KEGG enrichment analysis were performed on the inter-section targets using the DAVID tool.The top 20 KEGG pathways,core targets and drug components were used to construct a"component-target-path-way"network and the network visualization was performed using Cytoscape 3.9.1 software.The in-teractions between drug compounds and the tar-gets were evaluated by molecular docking,and the docking results were visualized using Discovery stu-dio.HFL-1 cells were cultured and the effect of the drug compounds on cell viability was determined by MTT assay.The inhibition rate was then calculat-ed to determine the optimal drug concentration.HFL-1 cells were cultured in vitro and were as-signed into 4 groups:control group,TGF-β1 group,TGF-β1+LD group(LD group),TGF-β1+HD group(HD group).CCK-8 kit was used to evaluate the anti-proliferative activity of the drug compounds against HFL-1 cells at 24,48 and 72 h.Plate clone formation assay was performed to evaluate the ef-fect of drugs on the colony formation ability of HFL-1 cells.RT-qPCR and western blot were conducted to determine the effect of the compounds on the mRNA and protein expression levels of α-smooth muscle actin(α-SMA),collagen type Ⅰ(COI-Ⅰ),and collagen type Ⅲ(COI-Ⅲ)in each group.RESULTS:A total of 197 intersection targets of Piperlongum L and anti-pulmonary fibrosis were identified.The core PPI network comprised 29 nodes(targets)and 199 edges(interactions).GO functional analysis showed that the significantly enriched biological processes associated with the compounds in Piper-longum L included negative regulation of apopto-sis,signal transduction,and protein phosphoryla-tion.Significantly enriched cellular components in-cluded cytoplasm,nuclear cytoplasm,plasma mem-brane.Enriched molecular functions associated with the compounds included the same protein binding,serine/threonine/tyrosine kinase activity,and protein binding.A total of 155 significantly en-riched KEGG signaling pathways were identified,with PI3K-Akt signaling pathway was highly associ-ated with PF and was the fourth most enriched pathway.PIK3CA,MAPK3,MAPK1,MTOR,SRC,CCND1,EGFR,PRKCA,BCL2,and GSK3B had the highest connectivity in the components-target-pathway network.Piperlongine,N-(2,5-dimethoxy-phenyl)-4-methoxybenzamide,tetrahydrotanshi-none,pisigenin and piperine were the key com-pounds in Piperlongum L.The molecular docking results showed that all the compounds except N-(2,5-dimethoxyphenyl)-4-methoxybenzamide had good binding activities with interactions observed with 10 proteins.The proliferation ability of the cells in the LD group was significantly lower than that of the TGF-β1 group at 48 h and 72 h(P＜0.05).The proliferation ability of cells HD group was sig-nificantly lower than the LD group at 24,48 and 72 h.The number of clones in each drug group was significantly reduced after treatment with the drugs(P＜0.05).The mRNA and protein expression levels of α-SMA,COI-Ⅰ,COI-Ⅲ in LD and HD groups were significantly lower than the expression levels in the TGF-β1 group.The protein expression levels of p-PI3K/PI3K and p-Akt/AKT were significantly lower in the two dose groups compared with the TGF-β1 group(P＜0.01).CONCLUSION:The results showed that the effect of Piperlongum L against PF is probably through modulation of the PI3K-Akt sig-naling pathway.

OBJECTIVETo identify the enterovirus from stool samples of patients with hand, foot and mouth disease(HFMD) in Guangzhou from 2010 to 2012 and to perform phylogenetic analysis of the VP1 gene sequences of coxsackievirus A4 and coxsackievirus A10.

METHODSA total of 5 484 samples of suspected cases of HFMD which Guangzhou Center for Disease Control received from 2010 to 2012 were collected.Virus RNA was tested by nested RT-PCR method as human enterovirus 71, coxsackievirus A16, coxsackievirus A4, coxsackievirus A10 and other enteroviruses positive, and 4 111 samples were positive. Phylogenetic tree was constructed by partial VP1 gene sequences of coxsackievirus A4 and coxsackievirus A10 to perform phylogenetic analysis.

RESULTSIn 4 111 enterovirus-positive samples, the positive rate of EV71, CoxA16, CoxA10 and CoxA4 was 35.1% (1 443/4 111) , 30.7% (1 261/4 111) , 2.0% (82/4 111),0.8% (31/4 111) respectively. Different enterovirus-positive rate was statistically significant (χ(2) = 148.34, P < 0.05) .Incidences of coxsackievirus A4 positive was highest in 3-year old children as 1.3% (7/534) , and that of coxsackievirus A10 positive was highest in 0-year old children as 3.7% (34/914) . The highest positive rate of diagnosed coxsackievirus A4 positive cases was admitted in April(2.6%, 12/460) , and the highest positive rate of diagnosed coxsackievirus A10 positive cases was admitted in August 4.3% (12/278). Phylogenetic analysis indicated that all the CoxA4 stains were divided into subtype A and subtype B, and the CoxA10 stains were divided into subtypes A, subtype B and subtype C. The VP1 gene nucleotide sequences of CoxA4 and CoxA10 this study measured both belonged to subtype A.

CONCLUSIONSThe VP1 gene nucleotide sequences of CoxA4 and CoxA10 in Guangzhou from 2010 to 2012 both belonged to subtype A.

Child ; China ; epidemiology ; Enterovirus A, Human ; Hand, Foot and Mouth Disease ; Humans ; Molecular Epidemiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral

Objective:To understand the epidemiological characteristics of imported COVID-19 cases in Guangzhou and provide scientific basis for the prevention and control of the disease.Methods:The data of imported COVID-19 in Guangzhou reported as of April 1, 2020 were collected from National Notifiable Disease Report System of China. The software Excel 2010 and SPSS 19.0 were applied for data cleaning and statistical analysis.Results:As of April 1, 2020, a total of 103 imported COVID-19 cases had been reported in Guangzhou, in which 92 were confirmed cases and 11 were asymptomatic infection cases. The number of the confirmed imported cases accounted for 11.4 % (92/806) in of the total in China at the same time. The male to female ratio of the cases was 1.58∶1 (63∶40). The median age of the cases was 31 years ( P 25- P 75:22-40 years), range of age was 11-63 years. The main occupational distributions of the cases were business services (41/103, 39.8 %) and students (36/103, 35.0 %). The imported cases whose destinations were 19 provinces and municipalities rather than Guangdong after entering the country accounted for 43.7 %. The main source countries of infections were the United Kingdom (27/103, 26.2 %), the Philippines (13/103, 12.6 %), the United States (13/103, 12.6 %) and Nigeria (7/103, 6.8 %). There were 34 inbound flights from which the imported COVID-19 cases were detected, in which 10 flights (10/34, 29.4 %) were found to carry more than 3 cases, with an average voyage time of (11.14±0.53) hours. A total of 29 imported cases(28.2 %) showed symptoms before entering the country, and 65 cases (63.1 %) had been isolated before the onset of the disease. The mean free activity time of the isolated cases after the onset was (6.76±0.79) days. The average number of the imported cases’ close contacts was 53. There were 13 clusters of COVID-19 caused by the imported cases, involving 36 cases (including 1 imported associated case). Conclusions:The sources of the imported COVID-19 cases in Guangzhou were widely distributed, and no cases had been found to be infected on the flights. In the early stage of the imported epidemic, there was high risk for the spread of the epidemic. Strengthened prevention and control of imported COVID-19 effectively reduced the of transmission risk of COVID-19 in communities.

Objective:To understand the epidemiological characteristics of mpox epidemic in Guangzhou and provide scientific evidence for the prevention and control of the disease.Methods:Based on the mpox surveillance system in Guangzhou, suspected mpox cases with fever and rash were reported by local hospitals at all levels to centers for disease control and prevention in Guangzhou for sampling, investigation and diagnosis. Descriptive epidemiological analysis was conducted on the clinical characteristics and treatment of the mpox cases and positive detection rate reported in Guangzhou as of 24:00 on June 23. Whole genome sequencing of the virus isolates was performed using Illumina Miniseq high-throughput sequencing platform.Results:The first mpox case in Guangzhou was reported on June 10 in 2023. As of 24:00 on June 23, a total of 25 confirmed mpox cases were reported. All the mpox cases were men with a M( Q1, Q3) of 32 (26, 36) years, the majority of the cases were MSM (96.0%). The main clinical features were rash (100.0%, 25/25), lymphadenectasis (100.0%, 25/25) and fever (52.0%, 13/25). Rash usually occurred near the genitals (88.0%, 22/25). The close contacts, mainly family members (40.4%, 23/57), showed no similar symptoms, such as fever or rash. The positive rate of mpox virus in household environment samples was 30.5%. The analyses on 3 complete gene sequences of mpox virus indicated that the strains belonged to West African type Ⅱb clade, B.1.3 lineage. Conclusions:Hidden transmission of mpox virus had occurred in MSM in Guangzhou. However, the size of affected population is relatively limited, and the possibility of wide spread of the virus is low.

Objective:To establish an index system of population based SARS-CoV-2 nucleic acid screening, and provide reference to determine the screening coverage appropriately.Methods:The literature review and brain storming sessions were used to develop the basic frame and index system of population based SARS-CoV-2 nucleic acid screening. Based on Delphi method and Analytic Hierarchy Process, 21 domestic experts were selected for two rounds of consultation to determine the index system of population based SARS-CoV-2 nucleic acid screening and its weight.Results:The positive indexes of experts in two rounds of consultations were both 100%. The experts' authority coefficients ( Cr) were 0.88±0.08 and 0.89±0.07, respectively. And the range of coefficient of variation ( CV) were (0.08, 0.24), (0.09, 0.25). The Kendall's W coordination coefficients were 0.34 and 0.22 respectively, which were statistically significant. The index system of population based SARS-CoV-2 nucleic acid screening was established, which had 4 first-level indexes, 11 second-level indexes and 58 third-level indexes. Besides, the weight of each index was determined. Conclusion:The index system of population based SARS-CoV-2 nucleic acid screening has been established, which can provide scientific reference for the health administration to determine the coverage of population based SARS-CoV-2 nucleic acid screening when local COVID-19 epidemic occurs.