Objective:To analyze the epidemiological characteristics and track probable imported sources of the local dengue outbreak in Zhangzhou city, Fujian province, 2019.Methods:Serum samples of patients with suspected dengue fever at acute phases were collected for virus detecting and serotyping by real-time fluorescence quantitative RT-PCR. For the positive specimens of local cases, full-length fragments of E gene were amplified by RT-PCR, and were sequenced and analyzed.Result:In 2019, there were 98 local cases of dengue fever in Zhangzhou city, which were concentrated in Zhao’an county, Longhai district and Yunxiao county. In this study, fourteen dengue virus E gene sequences representing different sources in different districts and counties were selected. The amino acid sequence virulence site analysis showed that the local epidemic strains were relatively virulent strains. The gene sequence alignment and phylogenetic analysis showed that all the local strains were classified as DENV-I subgenotype genotype I, divided into a, b and c three different branches. The evolutionary branch a contained all Zhao’an and Longhai sequences and was divided into three sub-branches, the b and c evolutionary branches were both the sequences of Yunxiao. There was a high correlation between the Shenqiao Town in Zhao’an and the Haicheng Town in Longhai. The other areas of the strains were limited to the towns, and the evolutionary branches were close to the other areas in China and countries in Southeast Asia.Conclusions:The indigenous dengue outbreaks in Zhangzhou, 2019 were caused by multiple sources of introduction and originated from other areas in China or from Southeast Asian countries and there was also the possibility of local cross-county transmission.

Objective:To analyze the epidemiologic characteristics of environmental samples of avian influenza virus in Fujian province from 2017 to 2021, and provide a reference for the prevention and control of avian influenza.Methods:Six types of specimens were collected from four types of environments in six cities in Fujian province. And the specimens were subjected to nucleic acid detection for influenza A, subtypes H5, H7 and H9 by fluorescence quantitative PCR, and the results were analyzed statistically with descriptive epidemiological methods.Results:From 2017 to 2021, a total of 4 214 samples were collected from 6 cities, of which the positive rate of avian influenza virus was 41.53%, and the positive rates of H5, H7 and H9 subtypes were 2.33%, 1.16% and 23.16%, respectively. The positive rate for mixed subtypes of H5 and H7 was 0.05%, the positive rate for mixed subtypes of H5 and H9 was 1.83%, the positive rate for mixed subtypes of H7 and H9 was 0.83%, the positive rate for mixed subtypes of H5, H7 and H9 was 0.09%, and the positive rate of A-type unclassified was 12.08%. The difference in avian influenza virus detection among different monitoring places ( χ2=517.57, P<0.001), different types of specimens( χ2= 51.58, P<0.001), and different cities ( χ2=458.34, P<0.001) was statistically significant. Among different monitoring places, the positive rate of avian influenza virus in urban and rural live poultry markets was the highest. The highest rate of positive detection was found in specimens from the cage surface, cleaning poultry sewage and poultry chopping board surface, with 48.09%, 47.34% and 45.66%, respectively. In terms of different cities, Sanming city had the highest positivity rate (56.00%), while Zhangzhou city had the lowest positivity rate(3.34%). And the positive rate was higher from November to February of the next year and June to August each year. Conclusions:The overall positive rate of avian influenza viruses in Fujian province was relatively high, with H9 subtype accounting for the main proportion. The monitoring of avian influenza viruses in winter, spring and summer should be strengthened. And effective measures should be taken to deal with avian influenza especially in urban and rural live poultry markets.

The incidence and mortality of HCC in China account for approximately 50% of all cases worldwide. Low early diagnosis rate and high postoperative recurrence rate are two major causes for poor 5-year survival rate of HCC patients in China. At present, multiple problems such as low performance and compliance of screening technology and lack of effective markers for predicting postoperative recurrence, remain to be resolved. Due to the simplicity and accuracy, new molecular markers, such as liquid biopsy, are expected to serve as supplementary tools to traditional screening and early warning approaches, thereby realizing early detection and accurate treatment of HCC. In this article, research progress upon the clinical application of liquid biopsy in early screening and prediction of postoperative recurrence of HCC was reviewed, and prospects the future research.
Biomarkers ; Carcinoma, Hepatocellular/pathology* ; Humans ; Liquid Biopsy ; Liver Neoplasms/pathology* ; Mass Screening ; Neoplasm Recurrence, Local

Objective:To develop a triplex real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay for dengue virus (DENV), yellow fever virus (YFV) and chikungunya virus (CHIKV), so as to achieve the rapid detection of these three viruses.Methods:The complete genome sequences of DENV(Ⅰ, Ⅱ, Ⅲ, Ⅳ), YFV and CHIKV were retrieved from Global Shared Database for comparative analysis, estimate its conservative region, specific primers and probes were designed, then a triplex real-time RT-PCR assay was developed. The specificity was evaluated by other viral nucleic acids. The sensitivity was evaluated by in vitro transcribed RNAs of DENV, YFV and CHIKV. The repeatability of the method was evaluated by independent repeated experiments with different concentrations of viral nucleic acids. DENV detection method was validated with dengue patient serum. YFV and CHIKV detection methods were validated with simulated positive samples. The sera from healthy people were used for negative validation. Results:This method has no cross-reaction with other viral nucleic acids. The limit of detection (LOD) of DENV (Ⅰ、Ⅱ、Ⅲ、Ⅳ), YFV and CHIKV in vitro transcribed RNAs were less than 21.55 copies/PCR, 21.25 copies/PCR, 21.85 copies/PCR, 22.75 copies/PCR, 22 copies/PCR, 45.65 copies/PCR. The standard deviation of Ct values of each concentration was less than 0.5 and the coefficient of variation was less than 3%. The positive rate of clinical and simulated positive samples was 100%, and the negative rate of healthy serum was 100%. Conclusions:A triplex real-time fluorescent quantitative RT-PCR assay for DENV, YFV and CHIKV detection was established, and proved to be specific, sensitive and repetitive.

Objective:To analyze the clinical efficacy and safety of camrelizumab combined with apatinib as a second-line therapy for unresectable hepatocellular carcinoma (HCC).Methods:Ninety-four cases with mid-and advanced-stage HCC who received camrelizumab combined with apatinib as second-line treatment were enrolled. Routine blood test, blood biochemical indexes, tumor stage, tumor imaging characteristics, previous treatment strategies and other clinical data before treatment were documented. Imaging examination follow-up results and adverse reactions during treatment were followed up until the end of follow-up or loss of follow-up or death. Kaplan-Meier method was used to analyze the clinical efficacy.Results:As of the last follow-up, 94 cases with mid-and advanced-stage HCC had received camrelizumab combined with apatinib as second-line treatment. Among them, 15 cases were lost to follow-up, 31 cases died, and 48 cases survived. The overall remission rate was 31.9%. The overall disease control rate was 71.3%. The median time to disease-free progression was 6.6 months. The median time to disease progression was not yet available. The 1-year cumulative survival rate was 62.3%. Grade 3 and above adverse reactions mainly included were thrombocytopenia (7.4%), abdominal pain (4.3%), active hepatitis (4.3%), leukopenia (4.3%), diarrhea (3.2%), hand-foot syndrome (3.2%). All adverse reactions were effectively controlled.Conclusion:Camrelizumab combined with apatinib can effectively prolong the survival period of patients with mid-and advanced-stage HCC, and it is well tolerated.

Objective:To isolate and identify the yellow fever virus (YFV) from the specimens of the imported yellow fever (YF) cases in Fujian province in 2016.Methods:Sixteen positive YFV nucleic acid samples including serum, urine and saliva were inoculated into C6/36 cells, respectively. The isolated strain was identified by YFV real-time RT-PCR. The complete gene sequence of this strain was obtained by high-throughput next-generation sequencing, and the phylogenetic tree was drawn.Results:Only one strain was isolated from the serum of one case three days after onset, and identified as a YFV strain by real-time RT-PCR. BLAST analysis showed that the complete gene sequence of this strain was identical to the strain CNYF01R/2016(KX268355) isolated from the first YF imported case in China in 2016. The phylogenetic tree showed that this strain belonged to the same phylogenetic branch as the epidemic strain Angola71 in Angola in 1971, and was significantly different from the 17D vaccine strain (X03700), indicating that the YFV strain isolated in this study belonged to the wild YFV strain of Angola genotype.Conclusions:An Angola genotype YFV strain was successfully isolated from samples of imported YF cases in Fujian Province in 2016.

Objective:We attempted to investigate the variation of amino acids and molecular evolution trend of hemagglutinin(HA) genes of H9N2 avian influenza virus (AIV).Methods:In this study, we collected and aligned HA sequences of H9N2 AIV that submitted before April 2020 in the EpiFlu database. Then we analyzed the amino acid differences in the key sites of HA gene in bioinformatics method, including receptor binding sites, cleavage sites and glycosylation sites.Results:84.92% sequences have glutamine(Q) at the HA226 site in place of leucine(L) 84.75% sequences from avian and 72.97% sequences from mammals contained such HA-Q226 L mutation. 96.26% sequences still maintained the characteristics of low pathogenic AIV at the cleavage site of HA gene, except that 180 sequences from avian that two basic amino acids were inserted. Introducing one or two potential glycosylation sites were observed in 58.68% HA sequences, while loss of glycosylation site at HA210-212 in 66.44% sequences.Conclusions:The key protein receptor binding sites of the HA gene of H9N2 AIV showed obvious trends to binding human receptor. It revealed H9N2 AIV′s increasing adaptation to mammals. The cleavage sites and glycoylation sites of HA protein have a mutation tendency of characteristics of highly pathogenic AIV.

Objective: To study the etiological characteristics of an imported Chikungunya fever (CHIK) epidemic in Fujian province in 2018. Methods: Serum samples collected at different days after the onset of the two CHIK cases were detected by real-time RT-PCR and ELISA. Structural protein E1 gene was amplified by RT-PCR and sequenced for nucleotide characteristics analysis and phylogenetic tree analysis. Results: RNA of Chikungunya virus (CHIKV) was detected in the 4 serum samples collected on the first 5 days of the disease, and the earliest IgM antibodies were detected in specimens on the 5th day of the disease, however, IgG antibodies were only detected in specimen on 10th day. Compared with the S27-African prototype strain, 12 mutant points were found in the amino acids of E1 genes in this study. The E1 genes of the two CHIK cases were exactly the same, and they were closest to the evolutionary relationship with the strain isolated in the Philippines in 2014. Their genotype was Asian genotype. Conclusions This epidemic was confirmed to have been imported from the Philippines after the infection with the Asian genotype CHIKV, which suggests that Fujian province should strengthen the monitoring of persons entering from the CHIK epidemic area, so as to prevent imported cases from causing local outbreaks.

Objective: To elucidate the epidemiological and etiological features of a local outbreak of dengue fever (DF) in Taijiang district in Fuzhou, Fujian province in 2017, and speculate possible viral source based on phylogenetic analysis. Methods: The clinical and demographic data of cases were collected through field investigation and the outbreak was characterized epidemiologically by descriptive method. The patient′s serum were collected and the adult mosquitoes were captured by anti-mosquito double-net method for the laboratorial test and viral isolation. The viral isolates were typed by real-time fluorescent RT-PCR and their full length of viral envelope (E) genes were amplified by RT-PCR and sequenced. The E gene sequences obtained in this study, together with the reference sequences, were used for the phylogenetic analysis. Results: A total of 13 cases of autochthonous DF were confirmed in the outbreak. All cases presented obvious clinical manifestations and clustered spatially and temporally. The Breteau Index (BI) of mosquito larva density was the highest in epidemic foci of Xingang street and was relatively low in surrounding areas. Four DENV-1 strains, three from patients and one from the captured adult Aedes albopictus, were isolated and identified by real-time RT-PCR. Full length E gene sequences of the four isolates were completely identical and phylogenetic analysis showed that the isolates were genetically closest to the strain (GenBank No. KT825033) from Vietnam in 2014, rather than the DENV-1 strains found in Fujian previously. Conclusions The DF outbreak occurred in Fuzhou in 2017 was caused by DENV-1 which was imported possibly from somewhere outside of Fujian province and subsequently led to local DF transmission in human via the mosquito Aedes albopictus.

Objective To evaluate the feasibility and accuracy of virtual navigation contrast enhanced sonography in accurately location of small liver tumors which cannot be detected by conventional sonography during microwave ablation therapy. Methods Twenty-three patients with 28 small liver tumors,which could not be detected by conventional sonography but CT/MRI,underwent microwave ablation with virtual navigation contrast enhanced sonography from January 2015 to March 2017 at Nanfang Hospital. After fusion of images from both sonography and CT/MRI,small liver tumors were ablated under the real-time monitoring of navigated sonography. Virtual navigation contrast enhanced sonography was also utilized to evaluate the efficacy of ablation after the ablation.All patients underwent CT/MRI examination at one month post-ablation to evaluate the efficacy of ablation. Results Virtual navigation system successfully provided image fusion for all patients and all lesions(image fusion efficacy was 100%). All patients underwent virtual navigation contrast enhanced sonography monitored microwave ablation.Only one patient received extra ablation since a small proportion of residual tumor checked after the initial ablation. No severe complications occurred in the present study. One-month after ablation,all patients showed complete ablation by further CT/MRI examinations.Conclusion Virtual navigation sonography can precisely target small liver tumors which are undetected by conventional sonography.Evaluation of lesions and ablation efficacy can be performed with the help of virtual navigation contrast enhanced sonography during the ablation period,which have shown satisfactory clinical efficacy.