Advances in research of interchangeable immunization with live attenuated Japanese encephalitis vaccines and inactivated vaccines / 中华预防医学杂志

Japanese encephalitis (JE) virus is the leading cause of vaccine-preventable encephalitis in Asia and the Western Pacific, which mainly invades central nervous system. Vaccination is the most important strategy to prevent JE. Currently, both live attenuated Japanese encephalitis vaccines (JE-L) and inactivated vaccines (JE-I) are in use. Due to the supply of vaccines and the personal choice of recipients, there will be a demand for interchangeable immunization of these two vaccines. However, relevant research is limited. By reviewing domestic and foreign research evidence, this article summarizes the current situation of the interchangeable use of JE-L and JE-I, and makes recommendations when the interchangeable immunization is in urgent need, so as to provide reference for practical vaccination and policymaking in China.

Comparson of the immunogenicity of genotypeⅠJapanese encephalitis virus subunit vaccine candidate antigens / 生物工程学报

To screen the best genotypeⅠJapanese encephalitis virus subunit vaccine candidate antigens, the prMEIII gene, the polytope gene and the prMEIII-polytope fusion gene of the GenotypeⅠJapanese encephalitis virus GS strain were cloned into prokaryotic expression vector pET-30a. The recombinant proteins were obtained after the induction and purification. The prepared recombinant proteins were immunized to mice, and the immunogenicity of the subunit vaccine candidate antigens was evaluated through monitoring the humoral immune response by ELISA, detecting the neutralizing antibody titer by plaque reduction neutralization test, and testing the cell-mediated immune response by lymphocyte proliferation assay and cytokine profiling. The recombinant proteins with the molecular weights of 35 (prMEIII), 28 (polytope antigen) and 57 kDa (prMEIII-polytope) induced strong humoral and cellular immune responses in mice. Compared with prMEIII-polytope and polytope proteins, the prMEIII protein induced a significant expression of IL-2 and IFN-γ (P0.05). The study suggests that the prMEIII protein can be used for the development of the Japanese encephalitis virus subunit vaccine.

Co-infection of Dirofilaria immitis and Japanese encephalitis virus in a spotted seal (Phoca largha) in the Republic of Korea

A 10-year-old male spotted seal presented with loss of appetite and decreased activity. Grossly, the internal organs revealed several filarial nematodes in the right ventricle of the heart and the pulmonary vessels. Histopathological examination of the brain revealed moderate nonsuppurative meningoencephalitis with glial nodules and neuronophagia. Japanese encephalitis virus (JEV) of genotype I was isolated from the brain. All nematodes were identified as Dirofilaria immitis. This is the first clinical case of co-infection with D. immitis and JEV in a seal, suggesting that the seal, may be a dead-end host, like the human and horse, for JEV.

Entomological investigation of Japanese encephalitis outbreak in Malkangiri district of Odisha state, India

BACKGROUND A severe outbreak of Japanese encephalitis (JE) and acute encephalitis syndrome (AES) with high case fatality was reported from Malkangiri district of Odisha state, India during September to November 2016 affecting 336 children with 103 deaths. OBJECTIVES The purpose of this study was to investigate the outbreak in the light of entomological determinants. METHODS Entomological investigation was carried out in 48 villages from four mostly affected Community Health Centres (CHCs) of Malkangiri district. Dusk collections of resting adults was done in villages from indoor and outdoor sites to record the density of mosquito species, including the known JE vectors, feeding behaviour, parity, dusk index and infection status with JE virus (JEV). FINDINGS The per man hour density and dusk index of JE vector species varied from 2.5 to 24.0 and 0.81 to 7.62, respectively in study villages. A total of 1136 mosquitoes belonging to six vector species were subjected to PCR and one pool of Culex vishnui was found to be positive for JEV. CONCLUSION The JE transmission in Malkangiri district was confirmed. Thorough screening of human blood samples of JE/AES suspected cases and JE vector mosquitoes for the presence of JEV during rainy season every year is recommended.

Serological and molecular epidemiology of Japanese encephalitis virus infections in swine herds in China, 2006–2012

Japanese encephalitis virus (JEV) is a mosquito-borne, zoonotic flavivirus causing viral encephalitis in humans and reproductive disorder in swine. JEV is prevalent throughout China in human; however, spatiotemporal analysis of JEV in Chinese swine herds has not been reported previously. Herein, we present serological and molecular epidemiological results and estimates of prevalence of JEV infections among swine herds in various regions of China. The results suggest that JEV infections are widespread and genotype I and III strains co-exist in the same regions. Therefore, there is an urgent need to monitor JEV infection status among swine herds in China.

Seroprevalence of Dengue Virus Antibody in Korea

PURPOSE: The number of dengue fever cases is rising due to increasing overseas travel. Vaccination makes severe dengue fever in seronegative individuals after vaccination when they exposure to wild-type dengue virus. We investigated the seroepidemiology of the dengue virus for monitoring of Korean dengue virus immunity and establishing the prevention of dengue infection. METHODS: The study was based on 446 residual sera collected from 98 infants (2 months to 1 year old), 152 adolescents (13 to 19 years old), 90 adults (20 to 50 years old), and 106 elderly participants (more than 65 years old) for other studies. Antibody levels for dengue virus immunoglobulin G (IgG) in each age group were measured using an enzyme-linked immunosorbent assay (ELISA). For each dengue virus IgG positive or equivocal result, an IgG ELISA was performed for Japanese encephalitis virus. RESULTS: Of the 446 serum samples, only 1 (0.2%) adolescent had a positive result from the dengue IgG antibody test. In the dengue virus IgG antibody test, 14 (3.1%) samples showed equivocal results (10 adolescents and 4 elderly). In the 1 positive case of dengue virus IgG, the Japanese encephalitis IgG test was also positive. In the 14 equivocal cases of dengue virus IgG, there were 6 positive, 3 equivocal, and 5 negative of Japanese encephalitis IgG. CONCLUSIONS: The seroprevalence rate of dengue virus was very low in Koreans. This study provides important data for establishing the policy for preventive measures of dengue fever. It will be necessary to continuously monitor for dengue virus immunity.

TaqMan Real-time RT-PCR Assay for Detecting and Differentiating Japanese Encephalitis Virus / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To detect Japanese encephalitis virus (JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction (RT-PCR) detection system was developed.</p><p><b>METHODS</b>By aligning the full-length sequences of JEV (G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay.</p><p><b>RESULTS</b>With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/µL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%.</p><p><b>CONCLUSION</b>A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes.</p>

Improvement of indirect enzyme-linked immunosorbent assay for detection of Japanese encephalitis virus antibodies in swine sera

Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT (PRNT₉₀). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and PRNT₉₀ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and PRNT₉₀ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and PRNT₉₀ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and PRNT₉₀ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

Generation of Japanese Encephalitis Virus-like Particle Vaccine and Preliminary Evaluation of Its Protective Efficiency / 病毒学报

The cDNA fragment of JEV prME gene was cloned into the baculovirus shuttle vector (bacmid) to construct a recombinant baculovirus vector, defined as AcBac-prME. Then the recombinant baculovirus Ac-prME was obtained by transfecting Sf9 cells with AcBac-prME. Western blot analysis and immunofluorescence results indicated that both prM and E proteins were efficiently expressed in Sf9 cells. Electron microscopy suggested that prME was assembled into JEV-VLPs. To further evaluate the potential of JEV-VLPs as vaccine, the mice were immunized with JEV-VLPs and then challenged with lethal JEV. The results of mice survival and pathological changes demonstrated that the JEV-VLPs performed complete protection against JEV-P3 strain and relieved pathological changes in the mice brain significant. This study suggest that JEV-VLPs would be a potential vaccine for Japanese encephalitis virus.

Establishment of a Multiplex RT-PCR for the Sensitive and Differential Detection of Japanese Encephalitis Virus Genotype 1 and 3

Japanese encephalitis (JE) is a zoonosis that affects the nervous system of humans and other animals. The genotype of JE virus (JEV) has shifted recently from genotype 3 (G3) to genotype 1 (G1) in Asia, including Korea. Thus, a rapid differential assay is required to make an accurate diagnosis of JEV genotype. In this study, we designed common and differential primer sets for JEV G1 and G3 to detect the JEV envelope (E) gene. The specific primer sets for JEV G1 and G3 specifically amplified the target gene. The detection limits of the three primer sets were 10(1.0), 10(2.0), and 10(2.0) TCID₅₀/reaction, respectively. No cross-reactivity was detected with non-JEV reference viruses. The multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay specifically differentiated JEV G1 from G3. Thus, a one-step multiplex RT-PCR assay was established to rapidly and differentially detect JEV. This assay will be useful for confirming JEV infections in animals and checking the JEV genotype in veterinary biological products.

Zika Virus Infection / 대한내과학회지

Zika virus was first isolated in from nonhuman primate in 1947. It is in the genus Flavivirus, closely related to other flavivirus like Dengue, West Nile, Yellow fever and Japanese encephalitis virus. Since 2007 epidemic in Yap island, zika virus infections had spread to the countries in Micronesia and South Pacific. In 2015, Zika virus outbreak occurred in Brazil and now more than 40 countries in American continents reported autochthonous infection. The virus is transmitted mainly by Ae. aegypti mosquito with many other Aedes mosquito species known as vector. Recently, Zika virus infection is known to cause severe neurological complications and congenital malformation. In this paper, we will review current knowledge on Zika virus history, biology, clinical characteristics and preventive method.

Comparison of the antigenic relationship between Japanese encephalitis virus genotypes 1 and 3

PURPOSE: The Japanese encephalitis virus (JEV) genotype circulating in Korea has changed from G3 to G1. Therefore, the purpose of this study was to compare the antigenic relationship between the two genotypes by using antibody tests. MATERIALS AND METHODS: Blood samples from 42 sows and 216 horses were collected, and their seroprevalence was monitored using the hemagglutination inhibition and virus neutralization tests. Antisera against JEV G1 and G3 were isolated and prepared from guinea pigs. The cross-reactivity of these two viruses was then compared using the neutralizing antibody test. RESULTS: We found that there was a difference in the seropositive ratios of JEV G1 and G3. However, the difference was dependent on the antibody test used. There was also an observed difference in the antigenicity between the two genotypes, as ascertained using the neutralizing antibody test. CONCLUSION: There is an evident difference in JEV antigenicity between the genotypes G1 and G3. Therefore, we propose monitoring of the seroprevalence of JEV, and reevaluating the antigenicity of the current vaccine by using the relevant tests.

A Centralized Report on Pediatric Japanese Encephalitis Cases from Beijing Children's Hospital, 2013 / 生物医学与环境科学（英文）

Fifteen pediatric cases of suspected Japanese encephalitis (JE) were reported in Beijing Children's Hospital during the late summer of 2013. The clinical manifestations in most cases included high fever, seizures, and abnormal magnetic resonance imaging findings. Twelve of 15 cases were laboratory-confirmed as JE cases by pathogen identification. Epidemiological investigations showed that five of the 12 laboratory-confirmed patients had an incomplete JE vaccination history. Follow-up investigations after discharge indicated that seven laboratory-confirmed JE patients without JE vaccinations had relatively poor prognoses, with an average Modified Rankin Scale (MRS) score of 2.6 when compared with the other five laboratory-confirmed, JE-vaccinated patients with an average MRS score of 0.5. The observation of pediatric JE cases among those with a history of JE vaccination warrants further attention.

Percepção dos graduandos de enfermagem sobre o seu envelhecimento / Percepción de los estudiantes de enfermería sobre lo proprio envejecimiento / Perception of nursing undergraduate students on self-aging

RESUMO Objetivo: analisar a percepção dos graduandos de enfermagem sobre o próprio envelhecimento. Método: pesquisa de abordagem qualitativa, realizada em agosto e setembro de 2011, com 18 graduandos de enfermagem de uma Universidade pública de Salvador (Bahia). Os depoimentos foram analisados por meio da Análise de Conteúdo. Resultados: apreendeu-se o núcleo temático: Percepção do graduando de enfermagem sobre o próprio envelhecimento e, a partir deste, emergiram duas subcategorias: A) O Não Pensar; B) O contexto influenciando no processo. Conclusão: os graduandos revelam que o envelhecimento está intrínseco ao desenvolvimento humano, e possui o vínculo familiar, a espiritualidade e atividade física como ferramentas fundamentais para um envelhecimento ativo. Entretanto, os mesmos relatam que, o modo de vida acelerado e estressante vivido na sociedade possibilita inserir hábitos considerados inadequados, como o consumo de “fast food” e álcool, que trazem influências negativas para o próprio processo de envelhecimento. .

RESUMEN Objetivo: analizar la percepción de los estudiantes de enfermería sobre su proprio envejecimiento. Método: estudio cualitativo, realizado en agosto y septiembre de 2011, con 18 estudiantes de enfermería de una universidad pública en Salvador/Bahia. Los datos fueron analizados através de análisis de contenido. Resultado: incautados el tema central: Percepción de alumnos de enfermería sobre su propio envejecimiento y de esto surgieron dos subcategorías: A) No creo; B) El contexto influye en el proceso. Conclusión: los estudiantes revelan que el envejecimiento es intrínseco al desarrollo humano, y tiene los vínculos familiares, la espiritualidad y la actividad física como herramienta clave para el envejecimiento activo. Sin embargo, el mismo informe que, debido a la forma de vida que se vive en la sociedad de ritmo rápido y estresante permite insertar hábitos considerados inadecuados, como el consumo de “comida rápida” y el alcohol y convertirse en influencias negativas para su propio proceso tuvo como objetivo analizar de los estudiantes de enfermería su propio envejecimiento. .

ABSTRACT Objective: to analyze the perceptions of nursing undergraduate students on their self-aging process. Method: qualitative study carried out between August and September, 2011 with 18 nursing undergraduate students of a public university in Salvador, Bahia. The interviews were analyzed by means of the Content Analysis method. Results: the following thematic concept was apprehended: Perceptions of nursing undergraduates on their self-aging, which generated two subcategories: A) The “don’t think about it” process; B) The context infl uencing the process. Conclusion: undergraduates reveal that the aging process is an intrinsic factor to human development. Family ties, spirituality and physical activity would be key mechanisms toward active aging. However, students also reported that their accelerated and stressed social lifestyles led to inadequate habits, such as the consumption of fast food and alcohol, which become negative infl uences in their aging process. .

Study on Spatial Dispersal and Migration Events of Japanese Encephalitis Virus / 病毒学报

To explore the spatial distribution mechanism of Japanese encephalitis virus (JEV), PhyML v3.0 was used to build phylogenetic tree using JEV sequences in the dataset. PAUP v4.0 and Migrapyhla softz ware were then used to analyze the migration events. The results showed that a total of 95 migration events were observed during the dispersal of JEV throughout Asia. Further analysis revealed that Thailand, and several Chinese provinces (including Shandong, Shanghai, Sichuan and Yunnan), were the main migration sources of JEV. JEV spread from these migration sources as follows: from Thailand to Australia, Cambodia, Tibet and India; from Shanghai to eastern coastal Asian regions and Yunnan; from Shandong to Korea, Zhejiang, Hubei, Shanxi and Liaoning; from Sichuan mainly to inland regions of China, as well as Vietnam and Japan; and from Yunnan to Zhejiang. This study indicated that frequent migration events occurred during the dispersal of JEV in the Asia and Pacific regions, and that Thailand, Shandong, Shanghai, Sichuan and Yunnan were the sources of JEV dispersal.

Detection of serum antibodies against Japanese encephalitis virus in bats in Hainan and Guangdong Provinces of China / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the prevalence of serum antibodies against Japanese encephalitis virus (JEV) in bats.</p><p><b>METHODS</b>Blood samples from the heart were obtained from bats captured in Guangdong and Hainan Provinces in 2013. The anti-JEV antibodies in bat sera were tested using indirect ELISA and virus neutralization test.</p><p><b>RESULTS</b>A total of 201 bat serum samples were tested, in which the total positivity rate of anti-JEV antibodies was 46.27% (93/201). The positive rate of anti-JEV antibodies in bats from Hainan and Guangdong Provinces was 88.89% (48/54) and 30.61% (45/147), respectively. All the samples from Rousettus leschenaultia, Miniopterus schreibersii, Pipistrellus abramus, and Rhinolophus macrotis were positive for anti-JEV antibodies, and up to 95.56% (43/45) of the samples from Miniopterus schreibersii (from Hainan Province) yielded positive results. Of the 28 samples with positive results by indirect ELISA, 15 showed positive results in virus neutralization test (53.57%) with neutralization antibody titers ranging from 1:10 to 1:28.22.</p><p><b>CONCLUSION</b>Bats from different regions and of different species can be naturally infected with JEV and have a high prevalence of anti-JEV antibodies in their sera. The role of bats in the natural cycle of JEV awaits further study.</p>

Etiology and molecular-epidemiological characteristics of viral meningoencephalitis of Zhejiang province in 2013 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate pathogens and molecular-epidemiology characteristics of viral meningoencephalitis in the monitoring sites of Zhejiang province, 2013.</p><p><b>METHODS</b>Cerebrospinal fluid and/or stool specimens were collected from suspected patients admitted to the monitoring hospitals in southern and northern Zhejiang province. Such specimen were subject to real-time qPCR for the detection of Human enterovirus (HEV), Japanese encephalitis virus (JEV), Mumps virus (MuV), Herpes simplex virus (HSV) and Cytomegalovirus (CMV). HEVs were isolated using the RD and Hep-2 cell lines, while VP1 genes from all HEV-positive isolates or RNA-positive specimen were amplified, sequenced, for homology and evolution analysis.</p><p><b>RESULTS</b>92 (38.5%) of the 239 samples collected from 229 patients were detected as virus nucleic acid positive, including 87 HEV positive samples, 1 MuV positive, 2 HSV positive, and 2 CMV positive; of the 87 HEV positive samples, 38 were further determined to be Coxsackievirus (CV) and 49 as Echovirus (E). 56 HEV strains were isolated from 239 (23.4%) samples. From the 31 cerebral fluid specimen of nucleic acid positive yet virus isolation negative, the most specimen were identified with E9 (9 specimen), followed by CVA9 (8 specimen); the viral serotype of Zhejiang province HEV were CVA9, CVB4, CVB5, E6, E7, E9, E11, E14, E16, E25 and E30, respectively. Predominant epidemic strains identified at southern and northern Zhejiang province were CVB5 and E6 respectively. The phylogenetic analysis of VP1 gene showed that all the HEV isolates in Zhejiang province were HEV-B.</p><p><b>CONCLUSION</b>The HEV-B was the main pathogen for viral meningoencephalitis in Zhejiang province in 2013, including 11 serotypes, while E7 was the first time to be isolated in Zhejiang province. The predominant isolates were CVB5 and E6 in southern and northern Zhejiang province respectively. The positive rate of viral nucleic acid detection was significantly higher than that of viral isolation. Regular EV isolation method was exposed to the risk of missing-detection of E9 and CVA9.</p>

Isolation and identification of mosquito-borne arboviruses in Yuncheng city, Shanxi province, 2012 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the species and distribution of mosquitoes and mosquito-borne arboviruses in Yuncheng city of Shanxi province, China.</p><p><b>METHODS</b>Mosquito samples were collected in 19 collection sites from Linyi county and Yongji city in Yuncheng city, in August, 2012. After identification and classification, all the specimens were homogenized and centrifuged to acquire supernatant before being inoculated to both C6/36 and BHK21 cells for viral isolation. Positive isolates were identified with arbovirus species-specific primers under RT-PCR, for further sequencing and phylogenetic analysis.</p><p><b>RESULTS</b>A total of 10 455 mosquitoes of 7 species in 4 genuese were collected. The predominant mosquito species in Linyi county was Culex pipens pallens (91.96%, 3 911/4 253), but the one in Yongji city was Culex tritaeniorhynchus (72.85%, 4 518/6 202). A total of 23 strains of viruses were isolated from the mosquito pools. 15 strains from Culex tritaeniorhynchus and Culex pipens pallens were identified as genotype I Japanese encephalitis virus (JEV). Four strains from Culex pipens pallens were identified as Culex flavivirus (CxFV). Three strains from Culex pipens pallens were identified as Culex pipiens pallens densovirus (CppDNV). One strain from Armigeres subalbatus and Aedes albopictus was identified as Getah virus (GETV).</p><p><b>CONCLUSION</b>Four kinds of arboviruses were isolated from the mosquito pools, including GETV and CxFV, which were isolated and documented in Shanxi province for the first time. In the city of Yuncheng, Culex tritaeniorhynchus had been the predominant species and major vector for transmitting JEV. Genotype I JEV remained the major JEV circulating in the local natural environment.</p>

Study on the genotypic characteristics of Japanese encephalitis virus and serology in the healthy population of Zhejiang province, 2012-2013 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the genotypic characteristics and the neutralizing antibody levels of Japanese encephalitis virus (JEV) and Japanese encephalitis (JE) in both vector mosquitoes and in healthy people of Zhejiang province.</p><p><b>METHODS</b>Virus was isolated from mosquitos sampled from the Monitoring Stations located in Xianju county during 2012 to 2013. Phylogenetic and homological studies were carried out on the E gene. A total of 1 263 blood specimens from 642 healthy people were collected before and after the seasons of JE epidemics. JEV neutralizing antibody was detected by the micro-neutralization test.</p><p><b>RESULTS</b>Twenty-five JEV strains were isolated from a total of 11 650 mosquitoes. The identity of nucleotide appeared as 87.8%-99.7% both from 2012 to 2013 and from 1982 to 2010 while as 87.7%-88.0% with vaccine strain SA14-14-2, in Zhejiang. The phylogeny tree of E gene indicated that the newly isolated virus belonged to genotype I but no mutation of amino acid sequence coding conformational epitope was identified in the envelop protein. Both positive rates and the geometric mean titer (GMT) of neutralizing antibody in healthy people were 31.5%-42.0% and 1 : 2.56-1 : 3.53 in Xianju county, during 2012 and 2013, respectively. Both of the two positive rates (χ(2)≤1.76, P > 0.05) and the two GMTs (u≤0.64, P > 0.5) for antibodies pre or post the epidemic season did not show significant differences.</p><p><b>CONCLUSION</b>JEV isolated in Xianju during 2012 and 2013 belonged to genotype I. The positive rates of JEV neutralizing antibody from healthy people in Xianju were less than 42.0%, which showed no significant differendes pre or post JE epidemic season.</p>

Research progress in mechanisms of cellular entry of Japanese encephalitis virus / 病毒学报

Japanese encephalitis virus (JEV) is a pathogenic mosquito-borne flavivirus which is responsible for outbreaks of severe viral encephalitis. The cellular entry of JEV is a prerequisite for Japanese encephalitis, so the understanding of its underlying mechanisms will provide more approaches for treating such disease. In recent years, increasing research has been conducted to investigate the mechanisms of cellular entry of JEV, and the results of research on other flavivirus have expanded the research directions for JEV. More methods will be used to suppress JEV infection because of the development of E protein antibodies and the discovery of several inhibitors of the cellular entry process. This review will summarize the recent advances in the mechanisms of JEV cellular entry and membrane fusion.