Epidemiology and genotypes analysis of human papillomavirus infection in Beijing, China.

BACKGROUND: This study aimed to investigate the epidemiology of high-risk human papillomavirus (HPV) in the female population in Beijing, China, and identify the relationship between HPV genotypes and host factors. METHODS: HPV testing was performed on women aged 15-89 (mean age 38.0 ± 10.9 years) from Beijing in 2020. High-risk HPV genotyping real-time polymerase chain reaction was used to determine HPV genotypes. The overall prevalence, age-specific prevalence, genotype distribution, and the correlation between HPV genotypes and cervical cytology were analyzed. RESULTS: Among the 25,344 study participants, the single and double infection rates were 18.8% (4,777/25,344) and 4.2% (1,072/25,344), respectively. A total of 6,119 HPV-positive individuals were found to have 91.6% negative results for intraepithelial lesion or malignancy (NILM), 5.8% atypical squamous cells of undetermined significance (ASC-US), 0.9% low-grade squamous intraepithelial lesion (LSIL), and 1.7% high-grade squamous intraepithelial lesion (HSIL). In single HPV infections, the HPV16 genotype was highly associated with cervical cytology severity (χ2 trend = 172.487, P < 0.001). Additionally, HPV infection rates increased gradually with age, and statistical differences were observed across age groups (χ2 = 180.575; P < 0.001). High-risk HPV genotypes were highly prevalent in women below 25 years of age and those aged 55-59 years. Cluster analysis revealed that the 13 HPV genotypes could be roughly divided into two groups in a single infection; however, patterns of infection consistent with biological characteristics were not observed. CONCLUSION: High-risk HPV was found in 24.1% of outpatients, with HPV52, HPV58, HPV16, HPV39, and HPV51 being the most common high-risk genotypes. Single high-risk HPV infection was predominant. HPV16, HPV39, HPV51, and HPV52 were associated with cervical lesion progression. HPV16 infection was especially worrying since it aggravates cervical lesions. Because the infection rates of the 13 HPV genotypes differed by age, the peak HPV infection rate should not guide vaccination, screening, and prevention programs. Instead, these initiatives should be tailored based on the regional HPV distribution characteristics. Moreover, it was determined that Beijing's populace needed to receive treatment for HPV39 infection.

Intranasal immunization with HRSV prefusion F protein and CpG adjuvant elicits robust protective effects in mice.

Human respiratory syncytial virus (HRSV) is a leading cause of lower respiratory tract infections in elderly individuals and young children/infants and can cause bronchiolitis and even death. There is no licensed HRSV vaccine. An ideal vaccine should induce high titers of neutralizing antibodies and a Th1-biased immune response. In this study, we used EXPI293 cells to express the fusion (F) protein with a prefusion conformation (PrF) and compared the safety and efficacy of intranasal immunization with PrF in combination with two mucosal adjuvants (CpG ODN and liposomes) in mice. After two intranasal administrations, mice in the PrF + CpG group produced high titers of neutralizing antibodies (4961) and a Th1-biased immune response compared with the PrF + Lipo group. The lung viral load of mice in the PrF + CpG group was significantly reduced (3.5 log) compared with that in the adjuvant control group, and the survival rate was 100 %, while the survival rate of mice in the PrF + Lipo group was only 67 %. At the same time, this immunization strategy reduced the pathological damage to the lungs in mice. In conclusion, the combination of PrF and CpG adjuvant is immunogenic, elicits a Th1 type immune response, and completely protects mice from a lethal HRSV challenge. It is worthy of further evaluation as an HRSV vaccine in clinical trials. Clinical trial registration. This study was not related to human participation or experimentation.

Genetic diversity of E6, E7 and the long control region in human papillomavirus type 16 variants in Beijing, China.

Background: High-risk human papillomavirus type 16 (HPV16) is a risk factor for cervical cancer. The progression from initial infection to cervical cancer has been linked to properties of the viral sequences. However, the distribution of HPV16 variants among Chinese women has not been extensively addressed and the role of HPV16 variants in the risk of cervical carcinogenesis remains poorly understood. Methods: HPV16 positive cervical exfoliated cell samples were collected from 249 women living in Beijing, China. PCR products from two fragments of E6-E7 and LCR of HPV16 in these samples were sequenced and analyzed. Results: Lineage A was found in the subjects, including A1, A2, A3 and A4 sublineages. Based on the HPV16 reference sequences, 26 nucleotide mutations of A4 sublineage and 39 nucleotide mutations of A1-3 sublineages were found in the E6, E7 and LCR of HPV16 isolates. Point mutations T843C, A7287C and A7872G of A4 sublineage were significantly associated with high-grade cervical lesions. The high-frequency sites in HPV16 LCR located at regions that can bind to multiple transcription factors. Conclusions: This study contributes to the identification of unique variants in the E6, E7 and LCR of HPV16 isolates infected in Chinese women. Mutations of T843C, A7287C and A7872G in A4 sublineages were significantly associated with high-grade cervical lesions, suggesting that mutations in the E7 and LCR region have potential effects on viral replication and progression of cervical cancer.

Upregulation of nuclear factor E2-related factor 2 (Nrf2) represses the replication of herpes simplex virus type 1.

BACKGROUND: Nuclear factor E2-related factor 2 (Nrf2) is an important transcription factor which plays a pivotal role in detoxifying reactive oxygen species (ROS) and has been more recently shown to regulate inflammatory and antiviral responses. However, the role of Nrf2 in Herpes Simplex Virus type 1 (HSV-1) infection is still unclear. In this study, the interaction between the Nrf2 and HSV-1 replication was investigated. METHODS: The levels of oxidative stress was monitored by using 8-hydroxy-2'-deoxyguanosine (8-OHdG) ELISA kits, and the dynamic changes of Nrf2-antioxidant response element (Nrf2-ARE) pathway were detected by Western Blot. The effect of Nrf2-ARE pathway on the regulation of HSV-1 proliferation was analyzed by Western Blot, Real-Time PCR and TCID50 assay. RESULTS: HSV-1 infection induced oxidative stress. Nrf2 was activated, accompanied by the increase of its down-stream antioxidant enzyme heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1) in the early stage of HSV-1 infection. The proliferation of HSV-1 was inhibited by overexpression of Nrf2 or treatment with its activator tert-Butylhydroquinone (tBHQ). On the contrary, silencing of Nrf2 promotes virus replication. HO-1 is involved in the regulation of IFN response, leading to efficient anti-HSV-1 effects. CONCLUSION: Our observations indicate that the Nrf2-ARE pathway activates a passive defensive response in the early stage of HSV-1 infection. Targeting the Nrf2 pathway demonstrates the potential for combating HSV-1 infection.

Exploring the dynamics and interplay of human papillomavirus and cervical tumorigenesis by integrating biological data into a mathematical model.

BACKGROUND: Cervical cancer is the fourth most common tumor in women worldwide, mostly resulting from high-risk human papillomavirus (HR-HPV) with persistent infection. RESULTS: The present discoveries are comprised of the following: (i) A total of 16.64% of the individuals were positive for HR-HPV infection, with 13.04% having a single HR-HPV type and 3.60% having multiple HR-HPV types. (ii) Cluster analysis showed that the infection rate trends of HPV31 and HPV33 in all infections as well as HPV33 and HPV35 in single infections in precancerous stages were very similar. (iii) The single/multiple infection proportions of HR-HPV demonstrated a trend that the multiple infections rates of HR-HPV increased as the disease developed. CONCLUSIONS: The HR-HPV prevalence in outpatients was 16.64%, and the predominant HR-HPV types in the study were HPV52, HPV58 and HPV16. HR-HPV subtypes with common biological properties had similar infection rate trends in precancerous stages. Especially, as the disease development of precancer evolved, defense against HPV infection broke, meanwhile, the potential of more HPV infection increased, which resulted in increase of multiple infections of HPV.

Hepatitis E outbreak in a mechanical factory in Qingdao City, China.

OBJECTIVE: In July 2018, recurrent hepatitis E cases were reported from a factory in Qingdao City, China. The aim of this study was to identify additional cases, and help prevent future incidents by identifying possible risk factors for infection. METHODS: Participants were asked to provide blood samples for hepatitis E virus (HEV) IgM and IgG antibodies screening, as well as liver function test. A questionnaire that assessed demographics, potential risk factors, and clinical symptoms was completed by participants. HEV RNA genotyping was performed using a nested Reverse Transcriptional Polymerase Chain Reaction (RT-PCR) method. Adjusted Poisson regression model for participant characteristics and risk factors was constructed for multivariate analysis. RESULTS: Overall, 41(14.5%, 41/283) participants had recent acute infection (21 of these were symptomatic). The result of multivariate analysis demonstrated a significant association of acute HEV infection with consumption of pig liver within the past two months (Relative Risk 2.61, 95% confidence interval (CI) 1.10-6.17, p=0.0294). Sequencing of HEV RNA from seventeen acute cases indicated three HEV isolates of genotype 4 induced this outbreak. CONCLUSIONS: This was probably a common-source foodborne hepatitis E outbreak, related to the consumption of undercooked pig liver.

Implementation of a study to examine the persistence of Ebola virus in the body fluids of Ebola virus disease survivors in Sierra Leone: Methodology and lessons learned.

BACKGROUND: The 2013-2016 West African Ebola virus disease epidemic was unprecedented in terms of the number of cases and survivors. Prior to this epidemic there was limited data available on the persistence of Ebola virus in survivors' body fluids and the potential risk of transmission, including sexual transmission. METHODOLOGY/PRINCIPAL FINDINGS: Given the urgent need to determine the persistence of Ebola virus in survivors' body fluids, an observational cohort study was designed and implemented during the epidemic response operation in Sierra Leone. This publication describes study implementation methodology and the key lessons learned. Challenges encountered during implementation included unforeseen duration of follow-up, complexity of interpreting and communicating laboratory results to survivors, and the urgency of translating research findings into public health practice. Strong community engagement helped rapidly implement the study during the epidemic. The study was conducted in two phases. The first phase was initiated within five months of initial protocol discussions and assessed persistence of Ebola virus in semen of 100 adult men. The second phase assessed the persistence of virus in multiple body fluids (semen or vaginal fluid, menstrual blood, breast milk, and urine, rectal fluid, sweat, saliva, tears), of 120 men and 120 women. CONCLUSION/SIGNIFICANCE: Data from this study informed national and global guidelines in real time and demonstrated the need to implement semen testing programs among Ebola virus disease survivors. The lessons learned and study tools developed accelerated the implementation of such programs in Ebola virus disease affected countries, and also informed studies examining persistence of Zika virus. Research is a vital component of the public health response to an epidemic of a poorly characterized disease. Adequate resources should be rapidly made available to answer critical research questions, in order to better inform response efforts.

Ebola RNA Persistence in Semen of Ebola Virus Disease Survivors - Final Report.

BACKGROUND: Ebola virus has been detected in the semen of men after their recovery from Ebola virus disease (EVD). We report the presence of Ebola virus RNA in semen in a cohort of survivors of EVD in Sierra Leone. METHODS: We enrolled a convenience sample of 220 adult male survivors of EVD in Sierra Leone, at various times after discharge from an Ebola treatment unit (ETU), in two phases (100 participants were in phase 1, and 120 in phase 2). Semen specimens obtained at baseline were tested by means of a quantitative reverse-transcriptase-polymerase-chain-reaction (RT-PCR) assay with the use of the target sequences of NP and VP40 (in phase 1) or NP and GP (in phase 2). This study did not evaluate directly the risk of sexual transmission of EVD. RESULTS: Of 210 participants who provided an initial semen specimen for analysis, 57 (27%) had positive results on quantitative RT-PCR. Ebola virus RNA was detected in the semen of all 7 men with a specimen obtained within 3 months after ETU discharge, in 26 of 42 (62%) with a specimen obtained at 4 to 6 months, in 15 of 60 (25%) with a specimen obtained at 7 to 9 months, in 4 of 26 (15%) with a specimen obtained at 10 to 12 months, in 4 of 38 (11%) with a specimen obtained at 13 to 15 months, in 1 of 25 (4%) with a specimen obtained at 16 to 18 months, and in no men with a specimen obtained at 19 months or later. Among the 46 participants with a positive result in phase 1, the median baseline cycle-threshold values (higher values indicate lower RNA values) for the NP and VP40 targets were lower within 3 months after ETU discharge (32.4 and 31.3, respectively; in 7 men) than at 4 to 6 months (34.3 and 33.1; in 25), at 7 to 9 months (37.4 and 36.6; in 13), and at 10 to 12 months (37.7 and 36.9; in 1). In phase 2, a total of 11 participants had positive results for NP and GP targets (samples obtained at 4.1 to 15.7 months after ETU discharge); cycle-threshold values ranged from 32.7 to 38.0 for NP and from 31.1 to 37.7 for GP. CONCLUSIONS: These data showed the long-term presence of Ebola virus RNA in semen and declining persistence with increasing time after ETU discharge. (Funded by the World Health Organization and others.).

Cervical clear cell adenocarcinoma infected with human papillomavirus type 18 in an adolescent.

BACKGROUND: Cervical clear cell carcinoma is one of the rare subtypes of cervical adenocarcinomas. Few cases of cervical clear cell carcinoma have been reported in adolescents. CASE: We present here a case of a 14-year-old adolescent female diagnosed with a stage II cervical clear cell carcinoma. The patient had no sexual history or diethylstilbestrol-exposure in utero. Polymerase chain reaction identified in the tumor the presence of human papillomavirus type 18, a high-risk genotype for cervical cancer development. The ovaries were retained during surgery and the patient was still alive without recurrence after 9 years. SUMMARY AND CONCLUSION: Positivity of HPV18 nucleic acids suggests an association between high risk HPV infection and cervical clear cell carcinoma in the case. Furthermore, in the treatment of young patients with cervical carcinoma, the risks associated with loss of ovarian function should be weighed against that of potential ovarian metastasis.

[Study for association between high risk human papillomavirus and cervical lesions in the samples from opportunistic screening].

OBJECTIVE: To investigate the association between HPV genotypes and cervical lesion in Hybrid Capture 2 (HC2) HPV test positive samples. METHODS: 602 cervical samples randomly obtained detected as HPV positive by the HC2 high-risk probe cocktail were determined by polymerase chain reaction (PCR)-reverse dot blot (RDB), among them 344 participated Thinprep Cytology test. RESULTS: 569 (94.5%) samples were successfully amplified. The most common HPV genotypes were HPV16 (31.6%), 52 (16.7%), 58 (15.1%), 56 (8.1%), 39 (7.9%); HPV16 or HPV16/18 infection was significantly related to ASCUS, LSIL and HSIL; 24 samples suggested single HPV infection with the genotypes not available for HC2. CONCLUSION: The most common HPV genotypes in random screening were HPV16, 52 and HPV58; Our data demonstrated the pseudo-positivity of HC2 test was 4.0%; HPV16 was a index for progression of HSIL.

Visual detection of high-risk human papillomavirus genotypes 16, 18, 45, 52, and 58 by loop-mediated isothermal amplification with hydroxynaphthol blue dye.

A simple, rapid, sensitive, qualitative, colorimetric loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB) was established to detect high-risk human papillomavirus (HPV) genotypes 16, 18, 45, 52, and 58. All initial validation studies with the control DNA proved to be type specific. The colorimetric type-specific LAMP assay could achieve a sensitivity of 10 to 100 copies at 63°C for 65 min, comparable to that of real-time PCR. In order to evaluate the reliability of HPV type-specific LAMP, the assay was further evaluated with HPV DNAs from a panel of 294 clinical specimens whose HPV status was previously determined with a novel one-step typing method with multiplex PCR. The tested panel comprised 108 HPV DNA-negative samples and 186 HPV-DNA-positive samples of 14 genotypes. The results showed that the sensitivity of HPV type-specific LAMP for HPV types 16, 18, 45, 52, and 58 was 100%, 100%, 100%, 100%, and 100%, respectively, and the specificity was 100%, 98.5%, 100%, 98.8%, and 99.2%, respectively, compared with a novel one-step typing method with multiplex PCR. No cross-reactivity with other HPV genotypes was observed. In conclusion, this qualitative and colorimetric LAMP assay has potential usefulness for the rapid screening of HPV genotype 16, 18, 45, 52, and 58 infections, especially in resource-limited hospitals or rural clinics of provincial and municipal regions in China.

[Study of human papillomavirus in biopsy tissue specimens of esophageal carcinomas in Linzhou city].

OBJECTIVE: To investigate the prevalence of human papillomavirus (HPV), particularly of high-risk HPV in biopsy tissue specimens of esophageal carcinomas in Linzhou city. METHODS: General nested primer sets were used to detected the whole HPV genotypes, following by HPV16 and 18 type specific PCR for the HPV16 and 18 detection respectively. RESULTS: All 18 biopsy samples were HPV positive, and HPV 16 was detected in 13 of the 18 samples, HPV 18 was detected in 4 of the 18 samples. CONCLUSION: The high rate of HPV in the esophageal carcinoma samples suggested that HPV infection may be an important etiologic factor in the development of esophageal cancer in Linzhou city.

[Study of human papillomavirus in small cell neuroendocrine carcinoma of the uterine cervix].

OBJECTIVE: To observe the infection of the high risk human papillomavirus (HPV) in the specimen of the small cell neuroendocrine carcinoma of the uterine cervix (SCNEC). METHODS: We extracted the nucleic acids in the formalin-fixed and paraffin-embedded specimen from a 33-year-old patient diagnosed as small cell neuroendocrine carcinoma of uterine cervix and detected the HPV genotype with the nested PCR. RESULTS: We identified HPV18, a high-risk genotype, in the specimen. CONCLUSION: The HPV detection with the nested PCR was available for identification of the HPV genotype(s) in the paraffin-embedded specimens of small cell neuroendocrine carcinoma of the uterine cervix with a high accuracy and sensitivity.

[Analysis of human papillomavirus type 61, 83 and 84 MY fragments and preparation of standard samples].

OBJECTIVE: To investigate the mutations in nucleotide and amino acid level in HPV61, 83 and 84 Shanxi isolates. METHODS: Amplified fragments of HPV61, 83 and 84 from human papillomavirus (Human papillomavirus, HPV) molecular epidemiologic survey of Shanxi Province using HPV consensus primers MY09/11 were cloned in pMD18-T vector, and the plasmids were sequenced, then nucleotide sequences and amino acid sequences were analyzed. RESULTS: HPV61 and HPV83 isolates were consistent with reference strains U31793 and AF151983 in nucleotide sequences; four mutations of nucleotide (C6760T, T6931C, T6951C and C6987A) were found in HPV84 isolate compared with reference strain AF293960, among which C6987A resulted in D441E and the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was higher than that of HPV83 and 84. CONCLUSION: HPV61 and HPV83 isolates were consistent with reference strains, four mutations of nucleotide and one mutation of amino acid were found in HPV84,the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was the highest among those three isolates.

[Human papillomavirus type 16 intratypic variant infection and risk for cervical neoplasia].

OBJECTIVE: To study the distribution of human papillomavirus type 16 (HPV16) variants and their clinical significance in Han women with Cervical Intraepithelial Neoplasia (CIN). METHODS: Randomly making a collection of DNA samples of cervical cells from 77 Han out-patients infected with HPV16, PCR amplification of HPV16 DNA fragments containing E6 and E7 genes and sequenced. To study the HPV16 variants types in these out-patients and explore the relationship between the HPV16 variants and CIN by comparing the E6 genes sequenced with the reference strains downloaded from the GenBank. RESULTS Among 77 patients, the minimum age is 21 years old, the maximum age is 56 years old, and the average age is 36.39 +/- 6.86 years old. 61 patients (accounting for 79.2%) were diagnosed as CIN II and higher grade lesions while 16 patients (accounting for 20.8%) as CIN I. In this research, only European variant and Asian variant were found by Parsimony analyses of the sequences. There are 38 Asian variants and 39 European variants. With Chi2 test, Chi2 = 0.0034, P = 0.9535 > 0.05, it suggested that there was no enough evidence to support Asian- and European-variants had the different risk in the cause of cervical intraepithelial neoplasia and cervical cancer. CONCLUSION: It was not found Asian- and European-variants of HPV16 had different effect on the cervical cancer, but found only two major variants-Asian- and European-variants in Han people in this research. So we have reason to speculate that there are two major HPV16 variants (Asian- and European-variants) in China's Han women, while other variants, especially high cancer-causing Asian/American variant are not common.

tRNA binds to cytochrome c and inhibits caspase activation.

The specific molecular events that characterize the intrinsic apoptosis pathway have been the subject of intense research due to the pathway's fundamental role in development, homeostasis, and cancer. This pathway is defined by the release of cytochrome c from mitochondria into the cytosol and subsequent binding of cytochrome c to the caspase activator Apaf-1. Here, we report that both mitochondrial and cytosolic transfer RNA (tRNA) bind to cytochrome c. This binding prevents cytochrome c interaction with Apaf-1, blocking Apaf-1 oligomerization and caspase activation. tRNA hydrolysis in living cells and cell lysates enhances apoptosis and caspase activation, whereas microinjection of tRNA into living cells blocks apoptosis. These findings suggest that tRNA, in addition to its well-established role in gene expression, may determine cellular responsiveness to apoptotic stimuli.

[Meta analysis on etiological relationship between human papillomavirus and esophageal carcinoma].

OBJECTIVE: To study the relationship between human papillomavirus (HPV) and esophageal cancer development in China. METHODS: We searched and collected the published articles in Chinese related to HPV and esophageal cancer, and selected the articles with the PCR approach to detect HPV in the esophageal cancer specimens. RESULTS: We filtered our publication collection with standards as (1) PCR as the detection approach, (2) specimens as the paraffin-embedded sections, and (3) description of the primer in the experiments, and fifteen articles were enrolled for our meta-analysis. Among the articles, totally 980 specimens were tested, and 460 were HPV positive with the average HPV prevalence was 46.9% (95% CI: 43.8%-50.0%), varied from 8.3%-69.8% in the different locations. On the other hands, among 556 specimens whose HPV detection spectrum included HPV16, 139 showed the positivity of HPV16, the average prevalence was 25.0%, (95% CI: 21.4%-28.6%) varied from 4.4%-63.4% dependent on the locations; among 485 specimens whose HPV detection spectrum included HPV18, thirty-three specimens showed the positivity of HPV18, the average prevalence was 6.8% (95% CI: 4.6%-9.0%) varied from 0%-19.0% dependent on the locations. Third, among the fifteen articles enrolled in the meta-analysis, four articles used the same primer set for HPV detection in totally 406 paraffin-embedded specimens with the prevalence of 40.2% (95% CI: 36.0%-45.4%) varied from 20.3%-67.6% in different locations. CONCLUSION: Our analysis result suggested the HPV prevalence in the esophageal cancer samples of China and clued the possible etiological relationships between HPV infection and the esophageal cancer development.

[Human papillomavirus detection in clear cell carcinoma of the cervix].

OBJECTIVE: To investigate the infection of the high risk human papillomavirus (HPV) in the specimen of the clear cell carcinoma of the cervix. METHODS: We extracted the nucleic acids in the formalin-fixed and paraffin-embedded specimens from a 37-year-old patient with clear cell carcinoma of the cervix and detected the HPV genotype with the nested PCR. RESULTS: We identified HPV18, a high-risk genotype, in the specimens. CONCLUSION: The HPV detection with the nested PCR was available for identification of the HPV genotype(s) in the paraffin-embedded specimens of clear cell carcinoma of the cervix (CCCC) with a high accuracy and sensitivity.