Ruiping Xu

Oral Human Papillomavirus Infection and Its Risk Factors among 5,410 Healthy Adults in China, 2009–2011

Background: Despite the established link between oral human papillomavirus (HPV) infection and a subset of oropharyngeal squamous cell carcinoma (OSCC), little is known about the epidemiology of oral HPV infection among healthy adults in China. Methods: Oral swab specimens and questionnaires were collected from 5,410 individuals (ages 25–65 years). HPV DNA in oral exfoliated cells was tested by general primer-mediated (SPF1/GP6+) PCR and sequencing. Univariate and multivariate analyses were performed to assess the associations between exposure factors and oral infection. Results: Alpha mucosal HPV types were detected in 0.67% [95% confidence interval (CI), 0.47%–0.93%] of 5,351 β-globin–positive specimens, and cutaneous HPV in 5.46% (95% CI, 4.86%–6.10%). HPV 16 and 3 were the most prevalent types of α mucosal (0.43%; 95% CI, 0.27%–0.64%) and cutaneous HPV (4.17%; 95% CI, 3.65%–4.74%), respectively. The prevalence of α mucosal HPV decreased with increasing age (25–65 years) from 0.93% to 0.36% (Ptrend = 0.033), and was associated with self-reported history of oral disease [adjusted odds ratio (OR), 4.78; 95% CI, 1.07–21.41]. In 1,614 heterosexual couples, cutaneous HPV in one partner was found to increase the other partners risk of cutaneous HPV infection (adjusted OR, 2.33; 95% CI, 1.22–4.48). Conclusions: Oral HPV infection, particularly with α mucosal types, is rare among healthy adults in China. A younger age and a history of oral disease imply higher risk of α mucosal HPV infection. Impact: This study addresses the paucity of epidemiological data on oral HPV infection among healthy population in China. Cancer Epidemiol Biomarkers Prev; 23(10); 2101–10. ©2014 AACR.

T cell receptor β-chain repertoire analysis reveals intratumour heterogeneity of tumour-infiltrating lymphocytes in oesophageal squamous cell carcinoma.

Oesophageal squamous cell carcinoma (ESCC) has a generally poor prognosis, due to the lack of effective treatment methods. Immunotherapeutic approaches based on tumour‐infiltrating lymphocytes (TILs) have demonstrated that durable responses are produced in some patients with solid tumours, which suggests the potential feasibility of clinical application of immunotherapy for ESCC. However, many of the basic characteristics of TILs in ESCC are poorly understood, including clonality, specificity and spatial heterogeneity of the response of TILs, which depends on the interaction between antigens and T cell receptors (TCRs). We used ultra‐deep sequencing of rearranged genes in TCR β‐chain (TCRβ) to profile the basic characteristics of T cells in tumour tissues (four to six regions from each tumour) as well as matched adjacent normal tissue and peripheral blood from seven patients diagnosed with primary ESCC. We found that T cell clones within ESCCs were quite different from those of the peripheral blood and even the adjacent normal tissues in general. Although there was a relatively higher degree of overlap of intratumoural TCRβ repertoires than those between the tumour and other tissues, intratumoural TCRβ repertoires were spatially heterogeneous. Due to the restricted sampling, high‐throughput TCRβ sequencing could characterize the diversity and composition of a limited (compartment‐dependent) fraction of the respective T cell clones in any individual ESCC, expanding our understanding of immune behaviour and immune response and shedding more light on ESCC immunotherapy. Copyright

Comprehensive mapping of the human papillomavirus (HPV) DNA integration sites in cervical carcinomas by HPV capture technology.

Integration of human papillomavirus (HPV) DNA into the host genome can be a driver mutation in cervical carcinoma. Identification of HPV integration at base resolution has been a longstanding technical challenge, largely due to sensitivity masking by HPV in episomes or concatenated forms. The aim was to enhance the understanding of the precise localization of HPV integration sites using an innovative strategy. Using HPV capture technology combined with next generation sequencing, HPV prevalence and the exact integration sites of the HPV DNA in 47 primary cervical cancer samples and 2 cell lines were investigated. A total of 117 unique HPV integration sites were identified, including HPV16 (n = 101), HPV18 (n = 7), and HPV58 (n = 9). We observed that the HPV16 integration sites were broadly located across the whole viral genome. In addition, either single or multiple integration events could occur frequently for HPV16, ranging from 1 to 19 per sample. The viral integration sites were distributed across almost all the chromosomes, except chromosome 22. All the cervical cancer cases harboring more than four HPV16 integration sites showed clinical diagnosis of stage III carcinoma. A significant enrichment of overlapping nucleotides shared between the human genome and HPV genome at integration breakpoints was observed, indicating that it may play an important role in the HPV integration process. The results expand on knowledge from previous findings on HPV16 and HPV18 integration sites and allow a better understanding of the molecular basis of the pathogenesis of cervical carcinoma.

Seroprevalence of Merkel cell polyomavirus in the general rural population of Anyang, China.

Background Despite the probably causal link between Merkel cell polyomavirus (MCPyV) infection and Merkel cell carcinoma (MCC), a rare but aggressive skin malignancy, little is known about the seroepidemiology of MCPyV among healthy adults in China. Methods Serum antibodies against MCPyV were evaluated by multiplex serology in a population-based study of 5548 adults (including 1587 heterosexual couples) aged 25–65 years who were enrolled from rural Anyang, China in 2007–2009. Univariate and multivariate logistic regression analyses were performed to assess the risk factors for the seropositivity of MCPyV. Results The seroprevalence for MCPyV was 61.0%. MCPyV seropositivity was significantly higher in males than in females (64.5% vs. 57.7%, P<0.001), and for both genders, showed a trend of increase with age (Male: P trend<0.001; Female: P trend<0.001). Furthermore, among antibody positives, antibody levels of MCPyV increased with advancing age (P trend = 0.017). MCPyV seropositivity of one spouse was significantly associated with that of the other partner (Adjusted OR = 1.32, 95% CI: 1.07–1.62). However, there was no association between sexual behaviors and the seropositivity of MCPyV. Conclusions High seroprevalence of MCPyV was observed in healthy Chinese individuals. Serological evidence suggests that nonsexual horizontal spread of MCPyV can occur among family members, and further research in this regard is needed.

Seroprevalence of 10 human papillomavirus types in the general rural population of Anyang, China: a cross-sectional study

Objectives Data on the seroprevalence of human papillomavirus (HPV) in China are limited. The objective of this study was to characterise the serological profiles of HPV infection in a rural Chinese population and help establish effective vaccine policy. Methods Serum antibodies against the major capsid protein L1 of 10 HPV types (HPV-3, 6, 11, 16, 18, 45, 52, 57, 58 and 75) were evaluated with Luminex-based multiplex serology in a population-based study of 5548 adults (including 1587 couples) aged 25–65 years enrolled from rural Anyang, China, in 2007–2009. Results The seroprevalence for any HPV type and any of the types HPV-6/11/16/18 was 64.8% and 34.4%, respectively. 30.3% of adults were seropositive for any mucosal high-risk (HR) HPV, and HPV-58 (10.6%), HPV-16 (9.7%) and HPV-18 (9.3%) were the three most common types. 24.8% of seropositive individuals were positive for multiple mucosal HR-HPV serotypes. Seroprevalence for most HPV types was similar among men and women. While mucosal low-risk HPV seropositivity was found to significantly decrease with age, the prevalence of antibodies to mucosal HR antigens showed a general trend of increase with age. The lifetime number of sex partners was independently associated with mucosal HR-HPV seropositivity. Positive correlation of spousal seropositivity was observed for mucosal HPV but not for cutaneous HPV. Conclusions HPV infection was common in both men and women in rural China. HPV seroprevalence differed significantly with age, sexual behaviour and spousal infection status. These findings will be useful for evaluating and establishing HPV vaccination programmes.

Concurrence of oral and genital human papillomavirus infection in healthy men: a population-based cross-sectional study in rural China

Human papillomavirus (HPV) infection, a primary cause of genital cancer, is also related to the increasing incidence of oropharyngeal cancer among young men. Relatively little is known about the concurrence of oral and genital infection among healthy individuals. Oral and genital swab exfoliated cells were collected simultaneously from 2566 men in rural China. Using general primer-mediated (SPF1/GP6+) PCR and sequencing, HPV testing results were obtained from 2228 men with both valid oral and genital specimens (β-globin-positive). The prevalence of HPV infection was 6.7% in the oral cavity and 16.9% for the external genitalia. Among 43 men (1.9%, 43/2228) with oral-genital coinfection, 60.5% (26/43) harbored an identical HPV type at both sites. The risk of oral HPV infection was higher among men with genital infection than among uninfected men (11.4% vs. 5.7%, Adjusted OR = 2.3, 95% CI: 1.6–3.4). In addition, having multiple lifetime sexual partners was a significant risk for oral-genital HPV coinfection (Adjusted OR = 2.6, 95% CI: 1.0–7.0; 2 partners vs. 1 partner). These findings provide a basis for further understanding the natural history and transmission dynamics of oral HPV infection.

The distribution and common amino acid polymorphisms of human papillomavirus (HPV)-31 variants in 2700 women from Northern China.

To investigate the distribution of Human papillomavirus (HPV)-31 A, B and C variants as well as the common amino acid polymorphisms in Chinese women, all 14 HPV-31 positive cervical exfoliated cell specimens identified from a descriptive study including ∼2700 women from Northern China were analyzed. HPV-31 positive specimens were identified by Mass Spectrometry and the fragments of partial Long Control Region, E6 and E7 were amplified and directly sequenced or cloned into vector and then sequenced to confirm the variant information. HPV-31 prevalence in Northern Chinese female population was 0.52%. Six different sequences represented all 14 isolates, and these isolates were subsequently classified into variant lineage A (9), B (0) and C (5) by phylogenetic analysis. Five common amino acid polymorphism sites (2 in E6 and 3 in E7) and a novel non-synonymous mutation were detected in the current study. Our investigation suggested that HPV-31 was much less detected in Chinese women population than that in western countries. A and C variants were commonly detected while B variants were rarely detected in this population.

An Old Story Retold: Loss of G1 Control Defines A Distinct Genomic Subtype of Esophageal Squamous Cell Carcinoma

Esophageal squamous cell carcinoma (ESCC) has a high mortality rate. To determine the molecular basis of ESCC development, this study sought to identify characteristic genome-wide alterations in ESCC, including exonic mutations and structural alterations. The clinical implications of these genetic alterations were also analyzed. Exome sequencing and verification were performed for nine pairs of ESCC and the matched blood samples, followed by validation with additional samples using Sanger sequencing. Whole-genome SNP arrays were employed to detect copy number alteration (CNA) and loss of heterozygosity (LOH) in 55 cases, including the nine ESCC samples subjected to exome sequencing. A total of 108 non-synonymous somatic mutations (NSSMs) in 102 genes were verified in nine patients. The chromatin modification process was found to be enriched in our gene ontology (GO) analysis. Tumor genomes with TP53 mutations were significantly more unstable than those without TP53 mutations. In terms of the landscape of genomic alterations, deletion of 9p21.3 covering CDKN2A/2B (30.9%), amplification of 11q13.3 covering CCND1 (30.9%), and TP53 point mutation (50.9%) occurred in two-thirds of the cases. These results suggest that the deregulation of the G1 phase during the cell cycle is a key event in ESCC. Furthermore, six minimal common regions were found to be significantly altered in ESCC samples and three of them, 9p21.3, 7p11.2, and 3p12.1, were associated with lymph node metastasis. With the high correlation of TP53 mutation and genomic instability in ESCC, the amplification of CCND1, the deletion of CDKN2A/2B, and the somatic mutation of TP53 appear to play pivotal roles via G1 deregulation and therefore helps to classify this cancer into different genomic subtypes. These findings provide clinical significance that could be useful in future molecular diagnoses and therapeutic targeting.

Epidemiology of self-rated health in rural China: a population-based cross-sectional study.

Self-rated health (SRH) has been shown to be a good predictor of mortality. Data on SRH and its associated factors in the Chinese general population are limited. This study aims to assess the epidemiology of SRH in rural Anyang, China. SRH (categorized as “healthy”, “fair” or “unhealthy”) was measured in a population-based study of 2,814 adults (including 697 couples) aged 25 to 69 who were recruited from rural Anyang in 2014. Of 2,814 subjects, 63.1% rated their health as “healthy”, whereas 28.1% and 8.8% rated their health as “fair” and “unhealthy”. Compared to males, females had a higher likelihood of reporting a better SRH. Health ratings declined with increasing age, unmarried status, lower education levels. Poor SRH was positively correlated with medical history as well as high levels of fasting plasma glucose and total cholesterol, but not with unhealthy lifestyle indicators including smoking, drinking, and obesity. High household income was predictive of better SRH in men but not in women. Among couples, a positive spousal SRH concordance was observed, although the strength of this concordance was low. These findings will be useful for formulation of appropriate strategies for improving risk perception and promoting general health in economically developing regions.

Incidence and clearance of oral human papillomavirus infection: A population-based cohort study in rural China

The natural history of oral human papillomavirus (HPV) infection which is linked with the increased incidence of oropharyngeal squamous cell cancer (OPSCC) has been incompletely studied. Oral swab specimens and questionnaire data were obtained bi-annually for up to 6 visits from 4314 healthy adults aged 25-69 in rural Anyang, China. HPV infection status was evaluated with PCR-based sequencing. Participants with at least two consecutive valid HPV results within the study period were included in the incidence and clearance analysis. Among 3289 participants included in this analysis (median follow-up time 18.3 months), incidence rates of mucosal HPV, oncogenic mucosal HPV and cutaneous HPV were 0.53 (95% CI: 0.39-0.73), 0.30 (95% CI: 0.20-0.46), and 4.17 (95% CI: 3.70-4.70) per 1,000 person-months respectively. Most newly acquired infections were cleared within one year. Recent practice of oral sex increased the risk of incident infection with mucosal HPV (Adjusted HR, 5.03; 95% CI, 1.16-21.73) and oncogenic mucosal HPV (Adjusted HR, 10.13; 95% CI, 2.14-48.06). Newly acquired oral mucosal HPV infections are rare and most are cleared within one year in rural Chinese. This study expands understanding of the natural history of oral HPV in countries with a lower incidence of HPV-OPSCC.The natural history of oral human papillomavirus (HPV) infection which is linked with the increased incidence of oropharyngeal squamous cell cancer (OPSCC) has been incompletely studied. Oral swab specimens and questionnaire data were obtained bi-annually for up to 6 visits from 4314 healthy adults aged 25-69 in rural Anyang, China. HPV infection status was evaluated with PCR-based sequencing. Participants with at least two consecutive valid HPV results within the study period were included in the incidence and clearance analysis. Among 3289 participants included in this analysis (median follow-up time 18.3 months), incidence rates of mucosal HPV, oncogenic mucosal HPV and cutaneous HPV were 0.53 (95% CI: 0.39-0.73), 0.30 (95% CI: 0.20-0.46), and 4.17 (95% CI: 3.70-4.70) per 1,000 person-months respectively. Most newly acquired infections were cleared within one year. Recent practice of oral sex increased the risk of incident infection with mucosal HPV (Adjusted HR, 5.03; 95% CI, 1.16-21.73) and oncogenic mucosal HPV (Adjusted HR, 10.13; 95% CI, 2.14-48.06). Newly acquired oral mucosal HPV infections are rare and most are cleared within one year in rural Chinese. This study expands understanding of the natural history of oral HPV in countries with a lower incidence of HPV-OPSCC.