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The in vitro transcribed (IVT) mRNA technology has progressed rapidly and the application of mRNA vaccines in the COVID-19 pandemic made it become the most talked-about topic. Compared with protein drugs, IVT mRNA has a lower cost; it can be modular produced and its sequence can be modified easily, so it has a broad application prospect. However, due to its short history, mRNA drugs face the problem of lacking sufficient clinical data, and there is no quality control standard for mRNA drugs except mRNA vaccines. We overview the sequence design, delivery vectors, administration, application prospect and safety considerations of mRNA drugs. We also discussed the quality control of mRNA drugs briefly.
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The AP2/ERF gene family is one of the largest transcription factor families in the plant kingdom, and plays an important role in response to biological and abiotic stresses, plant hormone responses, and plant growth and development. In this study, the AP2/ERF family of Panax notoginseng was identified by bioinformatics methods, and the physicochemical properties, structure, phylogenetic relationship, expression pattern and function of PnDREB4 gene of the family were analyzed. The results showed that 140 AP2/ERF family members were identified in P. notoginseng, which were divided into DREB, ERF, AP2, RAV and Sololit subgroups. The physicochemical properties and motifs of proteins were similar among the subgroups. There were 34 differentially expressed genes in the AP2/ERF family of Fusarium oxysporum infected P. notoginseng plants, and 19 genes were up-regulated. The expression level of PnDREB84 was up-regulated with the extension of Fusarium oxysporum infection time in the range of 0-96 h. The content of ABA and SA in P. notoginseng plants overexpressing PnDREB84 gene increased after 4 ℃ stress. The results showed that PnDREB84 gene plays a dual regulatory role in the process of biological stress and abiotic stress. PnDREB84 gene can be used as a potential molecular marker for the breeding of new varieties of P. notoginseng. The identification of AP2/ERF transcription factor and function analysis of PnDREB84 gene of P. notoginseng provided data support for the analysis of stress resistance mechanism of P. notoginseng and the breeding of new varieties.
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The bZIP (basic leucine zipper) gene family is one of the largest transcription factor families in eukaryotes, and its members play important roles in stress response, secondary metabolism, plant growth, seed development and other aspects. To investigate the biological functions of the bZIP (CsbZIP) gene in Cannabis sativa L., we systematically investigated the CsbZIP gene family using bioinformatics methods based on the whole-genome and transcriptome data. The results showed that 55 CsbZIP gene family members (CsbZIP1-CsbZIP55) were identified and distributed on 10 chromosomes, belonging to 12 subfamilies. The gene structure and protein motif distribution of the same subfamily members were similar. Segment repeats were the main reasons for the expansion of CsbZIP gene family. Cis-elements analysis showed that the promoter regions of 73 lipid synthesis genes contained G-box or A-box element. qRT-PCR showed that the relative expression levels of 7 CsbZIP genes and 7 lipid synthesis genes were relatively high in hemp seed. 7 CsbZIP genes had a significant positive correlation with 7 lipid synthesis genes. This study revealed the structural features, evolutionary patterns and expression patterns of CsbZIP, providing important clues for further study on the regulation of CsbZIP on oil metabolism of hemp seed.
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Patients exhibit good tolerance to messenger ribonucleic acid (mRNA) vaccines, and the choice of encoded molecules is flexible and diverse. These vaccines can be engineered to express full-length antigens containing multiple epitopes without major histocompatibility complex (MHC) restriction, are relatively easy to control and can be rapidly mass produced. In 2021, the U.S. Food and Drug Administration (FDA) approved the first mRNA-based coronavirus disease 2019 (COVID-19) vaccine produced by Pfizer and BioNTech, which has generated enthusiasm for mRNA vaccine research and development. Based on the above characteristics and the development of mRNA vaccines, mRNA cancer vaccines have become a research hotspot and have undergone rapid development, especially in the last five years. This review analyzes the advances in mRNA cancer vaccines from various perspectives, including the selection and expression of antigens/targets, the application of vectors and adjuvants, different administration routes, and preclinical evaluation, to reflect the trends and challenges associated with these vaccines.
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Objective To investigate interaction effect of hyperglycemia and hyperuricemia on the patients with abnormal alanine aminotransferase(ALT). Methods From March to November 2018, 5 223 cases with complete and suitable data were enrolled in the physical medical examination in Yichang, Hubei Province of China. The metabolic characteristics of the two groups (508 ALT anomaly cases and 513 normal cases) were compared and analysed, Logistic regression model was used to study the independent effects of risk factors, and the interaction between risk factors was analyzed by additive model and multiplicative model. Results Levels of uric acid, triglyceride, total cholesterol, low density lipoprotein-cholesterol, fasting blood glucose, systolic blood pressure, diastolic blood pressure, body mass index, aspartate aminotransferase were significantly higher than that of the control group(all P<0.05). After adjusting some confounding factors, multivariate Logistic regression analysis showed that risk of abnormal ALT was 5.62 times higher in subjects with hyperglycemia and hyperuricemia than in subjects without them(95% CI:1.65-19.73, P=0.004). Interaction analysis of risk factors for abnormal ALT showed that there was no multiplicative interaction between hyperglycemia and hyperuricemia, but with additive interaction, the synergy index was 3.02, the relative excess risk due to interaction was 3.09, the attributable proportion due to interaction was 54.98% and pure factor attribution interaction was 66.87%. Conclusions There are several abnormal metabolic indices in individuals with abnormal ALT. The positive interaction between hyperglycemia and hyperuricemia are among the important risk factors for abnormal ALT patients. They can significantly increase the risk of illness.
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The aim of this paper was to observe the combination therapy with total triterpenoids of Chaenomeles speciosa and omeprazole on indomethacin-induced gastric ulcer in rats, and explore its possible mechanism. Rats were randomly divided into normal group, model group, omeprazole monotherapy(3.6 mg·kg~(-1)) group, total triterpenoids of C. speciosa monotherapy(100 mg·kg~(-1)) group, total triterpenoids of C. speciosa and omeprazole combination therapy(100 mg·kg~(-1)+3.6 mg·kg~(-1)) group. Except for the normal group, the other groups were given indomethacin(20 mg·kg~(-1)) by oral once a day for 7 consecutive days. Then the treated groups were given corresponding drugs by gavage, once a day for 14 consecutive days. The next day after the last administration, half of the rats in each group were measured the gastric mucosal blood flow, gastric juice volume and serum TNF-α, IL-1β, IL-6, IL-4 and IL-10. After the remaining rats in each group were underwent pyloric ligation 4 hours after the last administration, the gastric endocrine volume, pH value and total acidity of gastric secretion were measured, then histological analysis was performed, MPO activity, cAMP content and histomorphological analysis were conducted. Real-time PCR was applied to detect the mRNA expressions of gastric tissue TNF-α,IL-1β, IL-6, IL-4, IL-10, VEGFA, A_(2A)R; the protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue were detected by Western blot. The results indicated that total triterpenoids of C. speciosa and omeprazole combination therapy might significantly increase gastric mucosal blood flow, gastric mucus volume, reduce gastric endocrine volume, secretion acidity and mucosal damage, decrease the levels of TNF-α,IL-1β and IL-6, increase the levels of IL-4 and IL-10 in blood and gastric tissue, inhibit the activity of MPO, increase the content of cAMP in gastric tissue, up-regulate the mRNA expressions of VEGFA, A_(2A)R and protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue, elevate p-PKA/PKA, p-CREB/CREB and p-EFGR/EFGR. Moreover, the combination therapy with total triterpenoids of C. speciosa and omeprazole was more obvious than those of two monotherapies. These aforementioned findings suggested that the combination therapy with total triterpenoids of C. speciosa and omeprazole on indomethacin-induced gastric ulcer have significant therapeutic effect on indomethacin induced gastric ulcer in rats, its mechanism might be related to regulating A_(2A)R/AKT/CREB, A_(2A)R/VEGFA, EGF/EGFR and MUC6/TFF2 signaling pathways, inhibiting pro-inflammatory factors, increasing gastric mucosal blood flow, up-regulating mucosal cell proliferation factors and promoting mucosal protective factors.
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Animals , Rats , Cytokines , Gastric Mucosa , Indomethacin , Omeprazole , Pharmacology , Phytochemicals , Pharmacology , Random Allocation , Rosaceae , Chemistry , Stomach Ulcer , Drug Therapy , Triterpenes , Pharmacology , Tumor Necrosis Factor-alphaABSTRACT
Objective To observe the effect of horticultural training based on healing garden as rehabilitation for post-operation of hand trauma.Methods From January,2015 to December,2016,78 patients with functional disorder after operation for hand trauma were random-ized into control group and observation group equally.All the patients accepted routine rehabilitation,while the observation group accepted horticultural training based on healing garden in addition,for 28 days.They were assessed with Total Active Movement(TAM),Disability of Arm Shoulder and Hand(DASH),and Upper Extremity Function Test(UEFT)before and after treatment.Results The scores of TAM, DASH and UEFT improved in both groups after treatment(t>10.632,P<0.001),and improved more in the observation group than in the con-trol group(t>2.269,P<0.05).Conclusion The horticultural training based on healing garden may promote the recovery of movement of fin-gers,function of hand and upper limbs in patients after hand trauma.
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Neovascular eye diseases are the leading cause of irreversible visual impairment in humans.Although intraocular anti-vascular endothelial growth factor therapy has become one of the most important treatments for these disease,the treatment itself is not stable and there are many potential risks and complications.So it is necessary to explore a clearer and more effective therapeutic target.Studies have shown that the inflammatory cytokines,especially TNF-α,IL-1,IL-6 and IL-18,are closely related to the onset of neovascular eye diseases.Therefore,a review on inflammatory cytokines in neovascular eye diseases will present in this article.
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Objective To establish a management system to promote early rehabilitation for work injured patients. Methods The 2015 hospitalized patients were set as control group (n=16,635), and the 2016 hospitalized patients were set as observation group (n=15,372). The control group was hospitalized for injury data collection surveys and was not implemented systematic rehabilitation propaganda, rehabilita-tion assessment and rehabilitation guidance. However, the observation group hospitalized after establishment of the Work Injuries Rehabilita-tion Investigation System and accepted work injuries rehabilitation intervention, such as rehabilitation propaganda, rehabilitation assessment and rehabilitation guidance. The changes of hand trauma rehabilitation indicators were compared between two groups. Results The rate of work injuries rehabilitation intervention was 80.6%in the observation group. In the observation group, the proportion of the patients who re-ceived early rehabilitation treatment in the designated hospital (56.8%) was significantly higher than that of the control group (17.0%) (χ2=2603.683, P<0.001);the proportion of injured patients who were admitted to the designated work injuries rehabilitation institutions (9.4%) was higher than that of the control group (4.3%) (χ2=285.906, P<0.001). The proportion of patients evaluated as disability in the observation group (17.4%) was significantly less than that of the control group (26.3%) (χ2=235.327, P<0.001). The disability score was significantly lower in the observation group (6.23±1.29) than in the control group (8.26±1.16) (t=68.371, P<0.001). Conclusion The Work Injuries Reha-bilitation Investigation System can promote the early rehabilitation for the hand trauma patients in the designated hospital for work injuries, improve the proportion of hand trauma patients to enter the designated work injuries rehabilitation institutions to receive rehabilitation treat-ment, reduce the proportion of disabled persons, and reduce the degree of disability, effectively promote the overall recovery of patients with hand trauma.
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Objective To establish a management system to promote early rehabilitation for work injured patients. Methods The 2015 hospitalized patients were set as control group (n=16,635), and the 2016 hospitalized patients were set as observation group (n=15,372). The control group was hospitalized for injury data collection surveys and was not implemented systematic rehabilitation propaganda, rehabilita-tion assessment and rehabilitation guidance. However, the observation group hospitalized after establishment of the Work Injuries Rehabilita-tion Investigation System and accepted work injuries rehabilitation intervention, such as rehabilitation propaganda, rehabilitation assessment and rehabilitation guidance. The changes of hand trauma rehabilitation indicators were compared between two groups. Results The rate of work injuries rehabilitation intervention was 80.6%in the observation group. In the observation group, the proportion of the patients who re-ceived early rehabilitation treatment in the designated hospital (56.8%) was significantly higher than that of the control group (17.0%) (χ2=2603.683, P<0.001);the proportion of injured patients who were admitted to the designated work injuries rehabilitation institutions (9.4%) was higher than that of the control group (4.3%) (χ2=285.906, P<0.001). The proportion of patients evaluated as disability in the observation group (17.4%) was significantly less than that of the control group (26.3%) (χ2=235.327, P<0.001). The disability score was significantly lower in the observation group (6.23±1.29) than in the control group (8.26±1.16) (t=68.371, P<0.001). Conclusion The Work Injuries Reha-bilitation Investigation System can promote the early rehabilitation for the hand trauma patients in the designated hospital for work injuries, improve the proportion of hand trauma patients to enter the designated work injuries rehabilitation institutions to receive rehabilitation treat-ment, reduce the proportion of disabled persons, and reduce the degree of disability, effectively promote the overall recovery of patients with hand trauma.
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AIM To study the programmed death effect of trametenolic acid B (TAB) against human gastric cancer HGC-27 cells.METHODS The proliferation of cells was examined by MTT assay;cell apoptosis rate and cell cycle were detected by flow cytometry;cell autophagy was observed by acridine orange staining and transmission electron microscope;expressions of autophagy-related proteins were detected by Western blot.RESULTS 10,20,40 μmol/L Trametenolic acid B had good growth-inhibitory effects on HGC-27 cells,blocked in G0/G1 phase,in a dose-dependent and time-dependent manner,but had no obvious effect on cell apoptosis.After treatment with 20 μmol/L trametenolic acid B for 24 h,a large number of autophagy vacuoles and autophagy bodies were observed under transmission electron microscope,and the proportion of cell autophagy was significantly increased with higher dose of TAB;TAB promoted the transformation of type Ⅰ microtubule-associated protein 1 light chain 3 (LC3 Ⅰ) to type Ⅱ microtubule-associated protein 1 light chain 3 (LC3 Ⅱ),up-regulated the expressions of myosin-like B-cell lymphoma-2 (Bcl-2) interacting protein (Beclin-1) and cyclin-dependent kinase inhibitor 1A (p21),and inhibited the CCND1 gene protein (CyclinD1) expression.CONCLUSION Trametenolic acid B can induce the autophagic death of HGC-27 cells,but it has no significant effect on cell apoptosis.
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Based on the trialistic theory of drug properties, substances, and efficacy in Tujia medicine, We adopted the one-way thinking and research method from efficacy to substance or vice versa and utilized the multicomponent and multi-target advantage of traditional medicine. At last, we had developed the bidirectional or multidirectional thinking and research method from efficacy to efficacy or from substance to substance. In the study, we used the backward selection method based on the known effects of current drug to fine another new role with known efficacy related by analyzing components, so as to find new efficacy or safer usage; We used the known substances as a starting points to find the new chemicals with the function compared to the known efficacy, thus the drug processing will be guided and the new medicinal resources and drugs will be found. The backward selection method is a simple and effective research method, which might make better use of traditional medicinal components and advantages of drug multi-target, and promote the rational use of traditional medicine and the research and development of new drugs.
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Bacillus subtilis is the focus of both academic and industrial research. Previous studies have reported a number of sequence variations in different B. subtilis strains. To uncover the genetic variation and evolutionary pressure in B. subtilis strains, we performed whole genome sequencing of two B. subtilis isolates, KM and CGMCC63528. Comparative genomic analyses of these two strains with other B. subtilis strains identified high sequence variations including large insertions, deletions and SNPs. Most SNPs in genes were synonymous and the average frequency of synonymous mutations was significantly higher than that of the non-synonymous mutations. Pan-genome analysis of B. subtilis strains showed that the core genome had lower dN/dS values than the accessory genome. Whole genome comparisons of these two isolates with other B. subtilis strains showed that strains in different subspecies have similar dN/dS values. Nucleotide diversity analysis showed that spizizenii subspecies have higher nucleotide diversity than subtilis subspecies. Our results indicate that genes in B. subtilis strains are under high purifying selection pressure. The evolutionary pressure in different subspecies of B. subtilis is complex.
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Bacillus subtilis , Genetics , Evolution, Molecular , Genes, Bacterial , Polymorphism, Single NucleotideABSTRACT
The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
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Amino Acid Sequence , Chromatography, High Pressure Liquid , Methods , Interferons , Reference Standards , Mass Spectrometry , Methods , Molecular Weight , Oxidation-Reduction , Peptide Mapping , Protein Processing, Post-Translational , Reference StandardsABSTRACT
This paper reports the determination of the drug antibody ratio in an antibody-drug conjugate with two methods, i.e. LC-MS and UV/VIS, and to provide a reliable method to scientifically evaluate and effectively control the drug antibody ratio. Deglycosylated sample was analyzed with C4 column followed by MS, and the number of conjugated drugs in the antibody was determined by the molecular weight increase due to the addition of different number of drugs to the antibody, and then drug antibody ratio was calculated by weighted average of different number of drugs conjugated to the antibody. Optical density at 252 and 280 nm was measured with UV/VIS, and due to the difference of extinction coefficients between the antibody and the drug, the drug antibody ratio was calculated from linear equation with two unknowns. The drug antibody ratio was 3.21 and 3.25 respectively measured by the two methods, and the results were similar with the two methods. Our study indicated that both methods, LC-MS and UV/VIS, could be applied to the analysis of drug antibody ratio of the antibody drug conjugate.
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Antibodies , Chemistry , Gas Chromatography-Mass Spectrometry , Methods , Glycosylation , Immunoconjugates , Chemistry , Maleimides , Chemistry , Molecular Weight , Pharmaceutical Preparations , Chemistry , Spectrophotometry, Ultraviolet , MethodsABSTRACT
The primary structure of TNK-tissue plasminogen activator (TNK-tPA) was characterized using liquid chromatography-mass spectrometry (LC-MS). Firstly, the molecular mass of deglycosylated protein was measured. Then peptide mass mapping and MS/MS of the reduced, alkylated and trypsin-digested sample were tested and analyzed so as to verify its amino acid sequence and identify post-translational modifications. Results show that the amino acid sequence was consistent with designed structure; about 5% of M207 was oxidized; T61 was fucosylated with -80% occupancy; N103, N448 and N184 (-15% occupancy) were glycosylated with complex-type oligosaccharides. LC-MS coupled with proper sample pretreatment is approved to be a rapid and powerful approach to characterize the primary structure of TNK-tPA.
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Amino Acid Sequence , Chromatography, Liquid , Glycosylation , Mass Spectrometry , Molecular Weight , Protein Processing, Post-Translational , Tissue Plasminogen Activator , ChemistryABSTRACT
The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.
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Amino Acid Sequence , Chromatography, High Pressure Liquid , Molecular Sequence Data , Peptide Mapping , Recombinant Fusion Proteins , Chemistry , Spectrometry, Mass, Electrospray Ionization , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tandem Mass Spectrometry , Vascular Endothelial Growth Factor A , ChemistryABSTRACT
Structure of a recombinant chimeric anti-CD20 IgG1 monoclonal antibody was verified by the application of high-performance liquid chromatography-mass spectrometry (HPLC-MS)and N-terminal sequencer. Molecular masses, N-terminal sequences and peptide maps of the antibody treated with different reagents and enzymes were measured. Results indicate that the amino acid sequences of light and heavy chains and 10 disulfide bonds were consistent with theoretical structure. By comparison of molecular masses and peptide maps for the fully glycosylated and deglycosylated samples, the N-linked glycosylation site was identified. The method is simple, rapid, precise, and could be referred to the quality control and structure determination of other IgG1 products.
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Amino Acid Sequence , Antibodies, Monoclonal , Chemistry , Antigens, CD20 , Allergy and Immunology , Chromatography, High Pressure Liquid , Glycosylation , Immunoglobulin G , Chemistry , Allergy and Immunology , Immunoglobulin Heavy Chains , Chemistry , Immunoglobulin Light Chains , Chemistry , Mass Spectrometry , Molecular Weight , Peptide Mapping , Recombinant Proteins , Chemistry , Trypsin , ChemistryABSTRACT
<p><b>BACKGROUND</b>Enterovirus 71 (EV71) and coxsackievirus A16 (Cox A16) are major causative agents for hand, foot and mouth disease (HFMD). Studies indicate that the frequent HFMD outbreaks result in a few hundreds children's death in China in recent years. The vaccine and other research for HFMD need to be developed urgently.</p><p><b>THE AIMS OF OUR STUDY WERE</b>to explore dynamic development of mother-source neutralizing antibodies against EV71 and Cox A16 in infants from Jiangsu Province, China, and to provide the fundamental data for further establishing of corresponding immunization course.</p><p><b>METHODS</b>Peripheral blood samples were collected from 133 of parturient women once immediately before delivery and their infants at two and seven months of age. Method of micro-dose cytopathogenic effect was used to measure neutralizing antibodies against EV71 and Cox A16, respectively.</p><p><b>RESULTS</b>Seropositive rates of anti-EV71 and anti-Cox A16 in prenatal women were 79.7% (106/133) and 92.5% (123/133), respectively; geometric mean titers (GMTs) were 29.0 and 61.9; 75.9% (101/133) prenatal women were both positive in anti-EV71 and anti-Cox A16; seropositive rates of anti-EV71 and anti-Cox A16 were 25.6% (34/133) and 38.3% (51/133) in infants at two months of age; GMTs were 12.3 and 18.0, respectively. GMTs of anti-EV71 were significantly higher for infants at seven months (82.6) compared with that at two months (P < 0.05), showing infants had inapparently infected by EV71 during two to seven months. Although only one offspring (0.75%) at seven months was found having anti-Cox A16 transfered from maternal, this observation suggested no maternal antibody may remain in infants at seven months.</p><p><b>CONCLUSIONS</b>The prevalence of EV71 and Cox A16 were relatively high in Jiangsu Province. Bivalent vaccine against both EV71 and Cox A16 should be developed, and the ideal time point for prime immunization for infants is around 2-5 months of age.</p>
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Female , Humans , Infant , Infant, Newborn , Antibodies, Neutralizing , Blood , Allergy and Immunology , Cells, Cultured , Enterovirus , Allergy and Immunology , Enterovirus A, Human , Allergy and Immunology , Hand, Foot and Mouth Disease , Allergy and Immunology , VirologyABSTRACT
Objective To investigate the characterization of the complete genome of EV71 in Beijing, 2008 and to provide basis for selecting appropriate virus strain to develop vaccine. Methods 12 throat swab samples were collected from children with hand-foot-mouth disease (HFMD). One sample named 08YM-3 was cultured and isolated in vero cells. Viral RNA was extracted and carried out by RT-PCR and 5' , 3' rapid amplification of eDNA ends (RACE) to obtain the sequence from 08YM-3. PCR products were cloned and analyzed. Nucleotide identity between sequences was calculated and sequence alignments were made to generate phylogenetic trees using MegAlign in DNAStar. Results 3 clones were constructed that covered EV71 complete genome. Data from sequences analysis showed that this viral strain named BJ08 shared 95.6%-96.7%, 88.3%-96.1%,78.1%-94.0%,90.8%-94.6%, 85.9%-94.1% and 90.9%-93.9% in 5' UTR, PI, P2, P3, 3' UTR region and complete genome with C4 subtype, respectively. B J08 showed low nucleotides identity (<90%) with other subtypes. Phylogenetic trees established from alignment of the complete genome and VPI region indicated that B J08 belonged to C4 subtype. BJ08 and C4 subtype strains shared the same amino acids in 6 sites in VP1 region, which were associated with EV71 subtype. There was no mutation in VP1 antigen epitope (92-107aa). Conclusion This BJ08 strain belonged to C4 subtype. Further study on EV71 complete genome would have great significance for vaccine research.