E2 Gene Research Articles

Ginkgo biloba extract mitigates the neurotoxicity of AlCl3 in alzheimer rat’s model: role of apolipoprotein E4 and clusterin genes in stimulating ROS generation and apoptosis

Purpose Alzheimer’s disease (AD) appears as a result of an increase in the accumulation of amyloid beta peptide (Aβ) and a decrease in neurotransmitters (acetylcholine) within the brain cells which may be due to increase in acetylcholinesterase (AchE) activity and change in expression of Apolipoprotein E4 (ApoE4) and Clusterin (Clu) genes. The aim of the present study was using natural products such as Ginkgo biloba (G. biloba) extract that has the potential to reduce Aβ formation and increase AchE inhibition with its ability to save neuronal DNA from damage. Methods Sixty male aged rats were divided into six experimental groups exposed to AlCl3 to induce AD model and were treated with G. biloba extract. Collected brain tissues were used to assess the apoptosis rate, reactive oxygen species (ROS) generation, AchE inhibitory activity, expression alteration in ApoE4 and Clu genes, DNA fragmentations and gutathione peroxidase (GPx) activity.Results: The results exhibited that rats exposed to AlCl3 increased significantly rate of apoptosis, ROS formation, DNA fragmentation, up-regulation of ApoE4 and Clu genes as well as decrease of AchE inhibitory activity and GPx activity compared with those in control rats. However, treatment of AlCl3-rats with G. biloba extract improved the above neurotoxicity results induced by AlCl3 exposure. Conclusions It is therefore likely that G. biloba extract’s protective properties against AD are due to its ability to activate the response against oxidative stress.

Phylogenetic and Mutation Analysis of the Venezuelan Equine Encephalitis Virus Sequence Isolated in Costa Rica from a Mare with Encephalitis.

Venezuelan Equine Encephalitis virus (VEEV) is an arboviral pathogen in tropical America that causes lethal encephalitis in horses and humans. VEEV is classified into six subtypes (I to VI). Subtype I viruses are divided into epizootic (IAB and IC) and endemic strains (ID and IE) that can produce outbreaks or sporadic diseases, respectively. The objective of this study was to reconstruct the phylogeny and the molecular clock of sequences of VEEV subtype I complex and identify mutations within sequences belonging to epizootic or enzootic subtypes focusing on a sequence isolated from a mare in Costa Rica. Bayesian phylogeny of the VEEV subtype I complex tree with 110 VEEV complete genomes was analyzed. Evidence of positive selection was evaluated with Datamonkey server algorithms. The putative effects of mutations on the 3D protein structure in the Costa Rica sequence were evaluated. The phylogenetic analysis showed that Subtype IE-VEEV diverged earlier than other subtypes, Costa Rican VEEV-IE ancestors came from Nicaragua in 1963 and Guatemala in 1907. Among the observed non-synonymous mutations, only 17 amino acids changed lateral chain groups. Fourteen mutations located in the NSP3, E1, and E2 genes are unique in this sequence, highlighting the importance of E1-E2 genes in VEEV evolution.

Identifying and elucidating the resistance of Staphylococcus aureus isolated from hospital environment to conventional disinfectants

BackgroundStaphylococcus aureus is a nosocomial pathogen, detection and elucidation of its resistance mechanisms to conventional disinfectants may aid in limiting its spread on environmental surfaces in health care settings. In the current study, disinfectant susceptibility of S. aureus strains isolated from the hospital environment as well as possible associations between the presence of disinfectant-resistance genes and reduced susceptibility to disinfectants was investigated. MethodsA total of 245 samples were collected from the hospital environmental surfaces. The minimum inhibitory (MIC) and bactericidal concentrations (MBC) of disinfectants against S. aureus isolates were determined using the micro-broth dilution method. The qac genes (qacA, qacE, and qacΔE1) were detected by PCR and confirmed by sanger sequencing. ResultsA total of 47 S. aureus strains were isolated, with more than 85% of them showing methicillin resistance. The qacA, qacE, and qac∆E1 genes were found in 23.4%, 29.7%, and 4.2% isolates respectively. All the isolates with qac genes had higher MIC and MBC values to selected disinfectants. ConclusionsSignificant methicillin resistant S. aureus (MRSA) contamination in the hospital environment was detected. Furthermore, higher qac gene frequencies were found in MRSA isolates that also correlated with higher MIC/MBC values to different disinfectants. The study proposes that hospitals should develop policies to determine disinfectant MICs against the common environmental isolates to contain the spread of resistant strains.

Oral squamous cell carcinoma in young people with human papilomavirus / Carcinoma de Células Escamosas oral em pessoas jovens com papilomavírus humano

Introduction: The growth of oral squamous cell carcinoma (SCC) in positive human papillomavirus (HPV) patients is observed. The epidemiological profile is: young men, non-smokers and non-alcoholics. This is attributed to changes in promiscuous sexual practice. Objective: To understand HPV carcinogenesis, epidemiological and preventive implications. Methods: An integrative review of the literature was carried out in March 2018 in the PubMed and Science Direct databases. The descriptors used were: SCC, Young, HPV and Staging, combined by the AND modulator. Inclusion criteria were articles published in the last 10 years, related to the thematic and in the English, Portuguese and Spanish languages. In the PubMed were found 75 articles, after the application of the inclusion criteria were selected 64 articles for study. In the Science Direct were found 8 articles, after applying the selection criteria was selected 1 article for analysis. Results: HPV subtypes 16 and 18 are the most virulent. HPV is carcinogenic through the E6 and E7 genes. E6 inactivates the tumor suppressor gene p53. E7 inactivates the tumor suppressor gene pRb, resulting in overexpression of the p16 tumor suppressor protein that is associated with a better prognosis. The impact of TNM staging changed over time, the effect of N staging on mortality reduced, while the impact of T staging increased. Conclusion: Protected sexual practice and vaccination are efficient measures for the control of SCC, HPV positive. Therefore, the campaigns should be extended to the male audience. In addition, the staging of the disease should be updated for a better prognosis.

Vaccine-induced thrombotic thrombocytopenia: Effect of E3 gene elimination from ds-DNA adenovirus vector?

Vaccine Induced Thrombotic Thrombocytopenia (VITT) is a very rare but serious adverse reaction to the Oxford–AstraZeneca and Johnson and Johnson (J&J) vaccines but not observed in the Pfizer and Moderna vaccines. It is immunologically and clinically almost identical to Heparin Induced Thrombocytopenia (HIT). However patients with VITT have no history of heparin exposure. The antigen provocateur has remained elusive. It has been suggested that it is vaccine DNA. However this does not necessarily explain why vaccine RNA is non-causative and why it is not reported as frequently in wild-type virus infection. Evidence suggests that it is the double stranded DNA specifically of the adenovirus vectors used in the Oxford-AstraZeneca and J&J vaccines; combined with the elimination of the E3 gene domain to promote immunity, that are responsible for VITT. We explain why the RNA vaccines are not implicated in the phenomenon. HIT is not exclusive to heparin but rather a patho-immunological response that can occur to others poly-anions of a particular charge and steric configuration. More appropriate terminology for HIT and VITT may be poly-anionic induced thrombotic thrombocytopenia (PITT).

Genetic variability of E6 and E7 genes of human papillomavirus type 58 in Jingzhou, Hubei Province of central China

BackgroundCervical cancer is a common malignant tumor in women, with a high mortality rate, has great harm to women’s health. Long-term and persistent infection of high-risk human papillomavirus (HR-HPV) is the main reason of the occurrence and development of cervical cancer.MethodsThe infection rate of HPV-58 is higher in the Jingzhou area. In this study, 172 complete HPV-58 E6-E7 sequences were amplified by polymerase chain reaction (PCR), the amplified products were sequenced, and the gene variations of HPV-58 E6-E7 were analyzed. A Neighbor-Joining phylogenetic tree was constructed by MEGA 11. The secondary structure of E6 and E7 protein was investigated. PAML X was used to analyze the selective pressure. The B cell epitopes of E6 and E7 proteins in HPV-58 were predicted by ABCpred server.ResultsIn E6 sequences, 10 single nucleotide variants were observed, including 7 synonymous and 3 non-synonymous variants. In E7 sequences, 12 single nucleotide variants were found, including 3 synonymous variants and 9 non-synonymous variants. There are 5 novel variants. The phylogenetic analysis showed that all the E6-E7 sequences were distributed in A lineage. No positively selected site was found in E6 sequence, but G63 in E7 sequences was identified as positively selected site. Some amino acid substitutions affected multiple B cell epitopes.ConclusionVarious E6 and E7 mutational data may prove useful for development of better diagnostic and vaccines for the region of Jingzhou, Hubei province of central China.

Genotypic diversity of CSFV field strains: A silent risk reduces vaccination efficacy of CSFV vaccines in Vietnam

Classical swine fever (CSF) is a highly contagious, devastating, and transboundary viral disease that afflicts swine industries worldwide. Immunization with vaccines is one of the most effective strategies for controlling this disease. However, shifts in the antigenicity and pathogenicity of novel evolving viral strains have the potential to evade vaccination. In this study, 352 samples from swines exhibiting fever, hemorrhages, lethargy, and diarrhea in different pig farms located in 9 provinces of Vietnam were collected. CSFV was identified even within farms that had been vaccinated against CSFV. Several farms had swine which had been co-infection with CSFV and other pathogens. Copies of the E2 gene of 21 samples were isolated, cloned, sequenced, analyzed, and compared with copies of E2 in four vaccine strains. We identified a total of 42 amino acid substitutions in this glycoprotein, including 11 positions that affect the antigenic properties of E2 and 7 positions that are associated with neutralizing epitopes. The E2 glycoprotein of CSFV strains circulating in Vietnam and vaccine strains differ in their antigenicity. These findings provide deep insights into the molecular characteristics, genetic diversity, pathogenicity, antigenicity, and evolution of CSFV strains in Vietnam. Understanding the pathogenicity, antigenicity, and evolution of circulating CSFV strains will provide avenues for developing new vaccines and efficient approaches to control this disease.

Rapid Differential Detection of Japanese Encephalitis Virus and Getah Virus in Pigs or Mosquitos by a Duplex TaqMan Real-Time RT-PCR Assay.

Both JEV (Japanese encephalitis virus) and GETV (Getah virus) pose huge threats to the safety of animals and public health. Pigs and mosquitoes play a primary role in JEV and GETV transmission. However, there is no way to quickly distinguish between JEV and GETV. In this study, we established a one-step duplex TaqMan RT-qPCR for rapid identification and detection of JEV and GETV. Primers and probes located in the NS1 gene of JEV and the E2 gene of GETV that could specifically distinguish JEV from GETV were selected for duplex TaqMan RT-qPCR. In duplex real-time RT-qPCR detection, the correlation coefficients (R2) of the two viruses were higher than 0.999. The RT-qPCR assay demonstrated high sensitivity, extreme specificity, and excellent repeatability. Detection of JEV and GETV in field mosquito and pig samples was 100 times and 10 times more sensitive than using traditional PCR, respectively. In addition, the new test took less time and could be completed in under an hour. Clinical sample testing revealed the prevalence of JEV and GETV in mosquitoes and pig herds in China. This complete duplex TaqMan RT-qPCR assay provided a fast, efficient, specific, and sensitive tool for the detection and differentiation of JEV and GETV.

Bioinformatics study of genes E1 and E2 from hepatitis C virus (HCV) with genotypes 1, 2, 3 and 6 as vaccine candidates for virus-like particles (VLPs)

Bioinformatics study of genes E1 and E2 from hepatitis C virus (HCV) with genotypes 1, 2, 3 and 6 as vaccine candidates for virus-like particles (VLPs)

Histone ubiquitination controls organ size in cotton (Gossypium hirsutum).

Ubiquitination plays a vital role in modifying protein activity and destiny. Ub-conjugating enzyme E2 is one of the enzymes that participates in this precise process. There are at least 169 E2 proteins in the allotetraploid cotton (Gossypium hirsutum), but their function remains unknown. Here we identify an E2 gene GhUBC2L and show its positive role in cell proliferation and expansion. Complete knock-down of GhUBC2L in cotton resulted in retarded growth and reduced organ size. Conversely, overexpression of GhUBC2L promoted cotton growth, generating enlarged organs in size. Monoubiquitination of H2A and H2B was strongly impaired in GhUBC2L-suppressed cotton but slightly enhanced in GhUBC2L-overexpressed plant. GhUbox8, a U-box type E3 ligase protein, was found to interact with GhUBC2L both in vivo and in vitro, indicating their synergistical function in protein ubiquitination. Furthermore, GhUbox8 was shown to interact with a series of histone proteins, including histone H2A and H2B, indicating its potential monoubiquitination on H2A and H2B. Expression of genes relating to cell cycle and organ development were altered when the expression of GhUBC2L was changed. Our results show that GhUBC2L modulates histone monoubiquitination synergistically with GhUbox8 to regulate the expression of genes involved in organ development and cell cycle, thus controlling organ size in cotton. This research provides new insights into the role of protein ubiquitination in organ size control. Histone monoubiquitination plays an important role in plant development. Here, we identified an E2 enzyme GhUBC2L that modulates histone monoubiquitination synergistically with an E3 ligase GhUbox8 to mediate organ size control in cotton.

The 'Why and How' of Cervical Cancers and Genital HPV Infection.

Knowing about the virology of human papillomavirus (HPV) including its structure, functions and mechanism of infection, helps in understanding the disease process and morphology of precancerous lesions for cervical cancer. Two types of HPV, low- and high-risk type, adopt different mechanisms of infection which cannot be differentiated on morphological basis. In addition to HPV infection, many other factors such as genetic predisposition, hormonal factors, host immune response, and multiple sexual partners can modify the course and progression of the disease. The viral genome comprises early and late proteins. These early and late genes are expressed in particular course of time after initial infection. Various products of early genes (E1–E7) coordinate for completion of viral life cycle in maturing squamous epithelium by utilizing replication factors and DNA polymerase enzyme of the host cell nucleus. The late genes are mainly concerned with packaging of the viral particles and their release through mature squamous cells. The episomal form of infection seen in the low-risk group of viruses results in productive infection whereas the integrated form seen in high-risk group of viruses is the basis of disruption of host cell growth cycle by inactivating two important tumor suppressor genes p53 and Rb gene by products of E6 and E7 genes. Cervical precancerous lesions and cancer are the resultant effect of the accumulation of mutations. This article discusses the virology of HPV, pathogenesis of HPV infection, and various other factors modifying the disease course.

Determination of Human Papillomavirus Type 18 Lineage of E6: A Population Study from Iran.

The epidemiological studies in Iran on HPV18 nucleotide changes are rare. This type of virus is prevalent in the Iranian population. Therefore, in the present study, we aimed to identify the genetic variability in HPV18 in the E6 region to evaluate the prevalence of lineage distribution and sublineages in a sample population in Iran. Overall, 60 HPV18 confirmed cases were investigated between 2019 and 2021. The specimens were collected, and molecular genotyping was done using the Linear Array HPV Genotyping Test. DNA extraction was performed by a viral DNA/RNA kit. The HPV E6 gene was amplified by using type-specific primers designed according to the HPV18 genome prototype sequence. The sequencing of the E6 region was successfully done on 43 samples which were then compared to the reference sequence. The most frequent sublineage of HPV18 in this study was A4 (69.7%), followed by A1 (18.6%) and A3 (11.6%). Neither A2 nor A5 sublineage was not detected in this study. The related nucleotide acid changes according to the main references were as follows: A3: T104C/T232G/T485C/C549A, A4: T104C/T485C/C549A. The predominance of A lineage with the high frequency of A4 sublineage was found in the present research. The importance of sublineages in susceptibility to a progressive form of infection requires to be more investigated among the different population.

Drug Sensitivity of Vaccine-Derived Rubella Viruses and Quasispecies Evolution in Granulomatous Lesions of Two Ataxia-Telangiectasia Patients Treated with Nitazoxanide.

A strong association between rubella virus (RuV) and chronic granulomas, in individuals with inborn errors of immunity, has been recently established. Both the RA27/3 vaccine and wild-type RuV strains were highly sensitive to a broad-spectrum antiviral drug, nitazoxanide (NTZ), in vitro. However, NTZ treatment, used as a salvage therapy, resulted in little or no improvements of RuV-associated cutaneous granulomas in patients. Here, we report investigations of possible causes of treatment failures in two ataxia-telangiectasia patients. Although a reduction in RuV RNA in skin lesions was detected by real-time RT-PCR, live immunodeficiency-related vaccine-derived rubella viruses (iVDRV) were recovered from granulomas, before and after the treatments. Tizoxanide, an active NTZ metabolite, inhibited replications of all iVDRVs in cultured A549 cells, but the 50% and 90% inhibitory concentrations were 10–40 times higher than those for the RA27/3 strain. There were no substantial differences in iVDRV sensitivities, neither before nor after treatments. Analysis of quasispecies in the E1 gene, a suspected NTZ target, showed no effect of NTZ treatments on quasispecies’ complexity in lesions. Thus, failures of NTZ therapies were likely due to low sensitivities of iVDRVs to the drug, and not related to the emergence of resistance, following long-term NTZ treatments.

Prevalence and distribution of human papillomavirus (HPV) in Luoyang city of Henan province during 2015\u20132021 and the genetic variability of HPV16 and 52

BackgroundPersistent high-risk Human papillomavirus (HPV) subtypes infection has been implicated as a causative of cervical cancer. Distribution and genotypes of HPV infection among females and their variations would assist in the formulation of preventive strategy for cervical cancer. The purpose of the present study is to investigate the prevalence of HPV among females in central China.MethodsThe distribution and genotypes of HPV among 9943 females attending the gynecological examinations in central of China during 2015–2021 were investigated. HPV genotypes were detected using a commercial kit. Nucleotides sequences of L1, E6 and E7 genes in HPV16 or HPV52 positive samples collected in 2021 were amplified by polymerase chain reaction (PCR). Variations of L1, E6 and E7 in HPV16 and HPV52 were gained by sequencing and compared with the reference sequence. Sublineages of HPV16 and HPV52 were determined by the construction of phylogenetic tree based on L1 gene.ResultsThe overall prevalence of HPV infection was 22.81%, with the infection rate of high-risk human papillomavirus (HR-HPV) was 19.02% and low-risk human papillomavirus (LR-HPV) was 6.40%. The most top five genotypes of HPV infection were HPV16 (7.49%), HPV52 (3.04%), HPV58 (2.36%), HPV18 (1.65%) and HPV51 (1.61%). Plots of the age-infection rate showed that the single HPV, multiple HPV, HR-HPV, LR-HPV infection revealed the same tendency with two peaks of HPV infection were observed among females aged ≤ 20 year-old and 60–65 year-old. The predominant sublineage of HPV16 was A1 and B2 for HPV52. For HPV16, The most prevalent mutations were T266A (27/27) and N181T (7/27) for L1, D32E for E6 and S63F for E7 in HPV16. For HPV52, all of the nucleotide changes were synonymous mutation in L1 (except L5S) and E7 genes. The K93R mutation was observed in most HPV52 E6 protein.ConclusionsThe present study provides basic information about the distribution, genotypes and variations of HPV among females population in Henan province, which would assist in the formulation of preventive strategies and improvements of diagnostic probe and vaccine for HPV in this region.

GmMDE genes bridge the maturity gene E1 and florigens in photoperiodic regulation of flowering in soybean.

Soybean (Glycine max) is highly sensitive to photoperiod, which affects flowering time and plant architecture and thus limits the distribution range of elite soybean cultivars. The major maturity gene E1 confers the most prominent effect on photoperiod sensitivity, but its downstream signaling pathway remains largely unknown. Here, we confirm that the encoded E1 protein is a transcriptional repressor. The expression of seven GmMDE genes (Glycine max MADS-box genes downregulated by E1) was suppressed when E1 was overexpressed and promoted when E1 was knocked out through clustered regularly-interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-mediated mutagenesis. These GmMDEs exhibited similar tissue specificity and expression patterns, including in response to photoperiod, E1 expression, and E1 genotype. E1 repressed GmMDE promoter activity. Results for two GmMDEs showed that E1 epigenetically silences their expression by directly binding to their promoters to increase H3K27me3 levels. The overexpression of GmMDE06 promoted flowering and post-flowering termination of stem growth. The late flowering phenotype of E1-overexpressing soybean lines was reversed by the overexpression of GmMDE06, placing GmMDE06 downstream of E1. The overexpression of GmMDE06 increased the expression of the soybean FLOWERING LOCUS T orthologs GmFT2a and GmFT5a, leading to feedback upregulation of GmMDE, indicating that GmMDE and GmFT2a/GmFT5a form a positive regulatory feedback loop promoting flowering. GmMDE06 also promoted post-flowering termination of stem growth by repressing the expression of the shoot identity gene Dt1. The E1-GmMDEs-GmFT2a/5a-Dt1 signaling pathway illustrates how soybean responds to photoperiod by modulating flowering time and post-flowering stem termination.

Identification of HPV16's E6 gene in suspected cases of cervical lesions and docking Study of its L1 protein with active components of Echinacea purpurae.

HPV 16 is the primary etiologic agent of cervical cancer and the presence of L1 and E6 oncoproteins are largely responsible for its virulence. It was the objective of this study to identify HPV16 isolates from suspected cases of cervical cancer at Specialist Hospital Sokoto and Sir Yahaya Memorail Hospiatal Birnin Kebbi, Nigeria and also to identify potent HPV16's L1 protein inhibitor using in silico analysis. A total of 144 cervical samples consisting of 21 low grade squamous intraepithelial lesion, 6 high grade lesion and 117 negative pap smears were collected. The samples were subjected for molecular detection using PCR targeting E6 gene of the virus. Data generated for the molecular prevalence was statistically analyzed using Chi-square method. AutoDock Vina was used to carry out the molecular docking between 2hr5 and Chicoric acid, curcumin and Echinacoside. Out of the 144 samples, 24 samples were positive for the PCR representing 16.9% molecular prevalence rate. There is statistically significant association between cyto-diagnoses and presence of HPV16 (P < 0.05). Docking analysis showed that the Chicoric acid components of Echinacea purpurae have strong binding affinity (-8.7 kcal/mol) to the L1 protein of the HPV. This study provides data on HPV 16 epidemiology in northern Nigeria, and also provides novel evidence for consideration on certain interacting residues, when synthesizing Anti-HPV compounds in the wet lab.

Year-round flowering gene e1, a mutation at the E1 locus on rice chromosome 7 and its combination with green revolution gene sd1 in an isogenic cell line.

Genetic analysis on the year-round flowering gene e1, which was derived from Kanto No. 79, an induced mutant by Koshihikari gamma-ray irradiated, was conducted through the backcross process to combine e1 and sd1 in the genetic background of Koshihikari. e1 strongly forwarded flowering 14 days earlier than the original variety Koshihikairi. Isogenic Koshihiakri combining e1 and sd1 was developed by four times of backcross with either Koshihikari or Koshihikari sd1 as recurrent parents by using the e1sd1 homozygous F3 plant in Koshihikari sd1 × Kanto No. 79 as a non-recurrent parent. As a result, "Koshihikari e1sd1" maturing 14 days earlier was approximately 30 cm shorter than Koshihikari. e1 was linked with DNA markers, which were near to Ghd7 on the short arm of chromosome 7. The whole genome sequencing revealed a single candidate SNP, which is specific to Koshihikari e1sd1, in the Xa21-like sequence, at 35213 bp downstream from Ghd7 on chromosome 7. Koshihikari e1sd1-specific SNP beside Ghd7 is expected to downregulate with Ghd7.

Construction and immune effect of an HPV16/18/58 trivalent therapeutic adenovirus vector vaccine

ObjectiveThis study aims to prepare candidate vaccines for cervical cancer immunotherapy by inserting the fused genes of human papillomavirus (HPV)16/18/58 mE6E7 lacking transforming activity into an adenovirus vector and to verify its efficiency in model mice with tumor expressing the associated HPV genes.MethodsThe E6/E7 genes of HPV16/18/58 were point-mutated to abolish their transforming activity, and adenovirus (AD)-HPV16/18/58 mE6E7 adenovirus vaccine was constructed. The immune effect of the adenovirus vaccine against HPV16/18/58-type tumors was analyzed by tumor morphology, enzyme linked immunosorbent assay, enzyme-linked immunospot and specific cytotoxic T lymphocyte (CTL) and T lymphocyte subsets.ResultsThe HPV16/18/58 mE6E7 plasmid containing point mutations was verified by quantitative real-time polymerase chain reaction (qRT-PCR), enzyme digestion and electrophoresis, and gene sequencing. qRT-PCR and Western blots verified that AD-HPV16/18/58 mE6E7 could express the HPV16 mE6E7, HPV18 mE6E7 and HPV58 mE6E7 fusion genes and proteins in cells. The results of animal experiments were as follows: In the vaccine group, the tumors formed later, the incubation period was longer, the growth was slower, growth was inhibited, and the survival period was significantly prolonged. The immunological results all showed that the vaccine could induce effective humoral and cellular immunity in mice with three types of tumors, compared with the phosphate buffered saline (PBS) group and the adenovirus-negative control (AD-NC) group, the differences were statistically significant (P < 0.05).ConclusionWe successfully constructed the HPV16/18/58 trivalent therapeutic adenovirus vaccine AD-HPV16/18/58 mE6E7. The AD-HPV16/18/58 mE6E7 adenovirus vaccine can protect immunized mice to a certain extent from TC-1, U14/LV-HPV18 E6E7 and U14/LV-HPV58 E6E7 cells, which contain HPV16, 18 and 58 E6 and/or E7 genes, respectively.

HPV16 E6 gene polymorphisms and the functions of the mutation site in cervical cancer among Uygur ethnic and Han nationality women in Xinjiang, China

BackgroundTo investigate the genotype distribution of human papillomavirus (HPV) in infected Uygur and Han women in Xinjiang, China; analyze the HPV16 E6 gene polymorphism site and relationship with the development of cervical cancer.MethodsThe HPV16 E6 sequence was analyzed using the European standard prototype to perform an evolutionary tree. HPV16 E6-T295/T350, G295/G350, and T295/G350 GV230 vectors were stably transfected into cervical cancer C33A cells to analyze the cell proliferation, migration and invasion, apoptosis by CCK8 and clonogenic assays, transwell and cell scratch assays, FACS experiments.ResultsThe total HPV infection rate was 26.390% (760/2879), whereas the Uygur 22.87% (196/857) and the Han was 27.89% (564/2022) (P < 0.05). Among 110 mutations, 65 cases of E6 genes were mutated at nucleotide 350 (T350G) with the leucine changing to valine (L83V). Moreover, there were 7 cases of E6 gene mutated at nucleotide 295 (T295G) with aspartic changing to glutamic (D64E). When E6 vector(s) of mutations sites were transfected into C33A cells, they were found to promote cellular proliferation, migration, invasion, and inhibit apoptosis. T295/G350-E6 was significantly stronger than G295/G350 and T295/T350, G295/G350 was significantly stronger than T295/T350 (P < 0.05). The T295/G350 had the strongest effect on C33A cells and G295/G350 was significantly stronger than T295/T350 (P < 0.05).ConclusionsThe positive HPV infection rates differed between the Uygur and Han in Xinjiang, China, and the genotype distribution of infection was different. After transfecting C33A cells with different eukaryotic expression vectors, the T295/G350 mutation site promoted the proliferation, migration, and invasion of C33A cells to a greater extent than G295/G350; however, G295/G350 had a stronger effect than T295/T350.

Genomics of Human Papillomavirus-induced Cervical Cancer

Human papillomavirus (HPV) induced cervical cancer is a serious health issue among most women from the least developed countries of the world due to scarcity of resources. HPVs have evolved a masterly infectious cycle that grabs the advantage of the self-renewal property of stratified cutaneous and mucosal epithelia. Firstly, the viral genome replicates episomally at a low copy number in the epithelial basal layers cells, with minimum viral transcription and translation. After that, when the infected cells are transmitted through the differentiation process maximum viral DNA synthesis and gene expression occur. The two major oncoproteins - E6 and E7, are responsible for inactivating the important tumor suppressor proteins, retinoblastoma (pRb) and p53. Due to the inactivation of these proteins, disruption occurs in the DNA replication, DNA repair mechanisms, oxidative induced damage (8-oxoguanine), aneuploidy, and apoptosis, leading to tumorigenesis. Hence, manipulation of E6 and E7 genes shows a successful result in the treatment of cervical cancer. The diagnosis tests include liquid-based preparations to improve the standard of the Pap smear; computer-based screening methods to improve Pap smear interpretation; and HPV testing methods that may be beneficial in triaging patients with untypical squamous cells of unknown significance or low grade squamous intraepithelial lesions (SILs). The continued studies of the molecular biology of HPVs are necessary to develop advanced screening techniques for testing and prophylactic vaccines for the elimination of HPV infection, also better therapeutic vaccines for the treatment of various HPVs infections.