This study aimed to evaluate the performance of two different principles of HPV testing in primary cervical cancer screening and ASC-US triage in rural areas.3,328 and 3,913 women were enrolled in Shanxi, China in 2017 and 2018, respectively, and screened using liquid-based cytology and different HPV tests with a 4-year follow-up. Different screening methods commonly used in clinical practice were evaluated.p < 0.001). Fifty-three cases of CIN2+ were accumulated over 4 years. The HPV infection rate in the HPV DNA chip test cohort was 21.10%. A total of 26 CIN2+ cases were identified at baseline, all in the HPV-positive population . 54 CIN2+ cases were cumulative over 4 years. At 4-year follow-up, HPV-negative results were significantly more protective against cervical intraepithelial neoplasia grade 2 or worse (CIN2+) than normal cytologic results at baseline. HPV screening was more sensitive and specific than cytologic screening (using ASC-US as the threshold) and performed better on the HPV DNA microarray test. In addition, compared with HPV 16/18 testing, sensitivity increases and specificity decreases when using HPV testing for cytologic ASC-US triage, regardless of which HPV test is used.In the HPV PCR test cohort, the prevalence of HPV infection was 14.90%. A total of 38 cases of CIN2+ were identified at baseline, 2 of which were in the HPV-negative cohort and the rest in the HPV-positive cohort (2 = 186.85, In the rural areas where we implemented the study, HPV tests performed well for screening than LBC and HPV DNA chip testing performed better than HPV PCR testing in the screening cohort. Optimal screening was achieved technically when used in combination with LBC for ASC-US population triage, without thinking the feasibility for resource availability. Cervical cancer is one of the top three most common malignancies among women worldwide. The Global Burden of Cancer 2020 report published by the International Agency for Research on Cancer of the World Health Organization showed that there were about 604,000 new cases of cervical cancer and 342,000 deaths worldwide in 2020; and there were about 110,000 new cases and 59,000 deaths in China, accounting for 18.2 and 17.3% of the global incidence and deaths, respectively. In addition, there was a trend toward younger age . PersistThe main kit technologies that have been marketed for HPV nucleic acid detection are the fluorescent PCR method, Hybrid Capture 2 (HC2), biochip, and flow-through fluorescent hybridization . There iTwo counties of Changzhi City, Shanxi Province (Changzhi county and Wuxiang County) were selected to establish cervical cancer screening cohorts. From May to June 2017, 3,328 rural women in Changzhi county underwent cervical cancer screening; From August to September 2018, 3,913 rural women in Wuxiang County underwent cervical cancer screening.The inclusion criteria were as follows: (1) age ≥ 21 years with an intact uterus; (2) no history of cervical cancer treatment or cervical surgery; (3) no sexual intercourse within 48 h after screening and no vaginal medications, vaginal contraceptives, or vaginal washings within 48 h; (4) no suspected clinical pregnancy symptoms; pregnant women could participate in the study up to 8 weeks after the end of pregnancy; (5) signed informed consent.All study participants were included in the study after giving informed consent, completing a questionnaire and a gynecological examination. The questionnaires were administered by trained investigators and included general information about the study participants, such as marital status, education level, history of smoking and alcohol consumption, menstrual history and reproductive history.A trained gynecologist performed gynecological examinations of the vulva, vagina, and cervix for each enrolled woman. Participants in Changzhi County had one cervical exfoliated cell collected by a gynecologist and preserved in cytology preservation solution (PreservCyt solution) for liquid-based cytology testing (LBC) and HPV DNA testing. Participants in Wuxiang County had two cervical samples collected by gynecologists: one cervical exfoliated cell was collected using a single-use cervical sampling swab and preserved in cytology preservation solution (PreservCyt solution) for LBC testing; one cervical sample  was collected and preserved in cell preservation solution  in a single-use sampling kit for HPV DNA testing.Within 12 weeks of collection of cervical exfoliated cell specimens, those with positive results for HPV types 16 and/or 18 and cytology results ≥ASC-US were referred to undertake colposcopy by an experienced gynecologist. If the colposcopy result was abnormal, a cervical biopsy or endocervical curettage (ECC) was performed on the subject. Participants with HPV-negative test results and normal cytology results at baseline were followed up for cervical cytology after 3 years. In contrast, participants with HPV-positive test results or cytology ≥ASC-US at baseline received annual cervical cytology follow-up (3 years). Participants with ≥ASC-US results at follow-up received colposcopy, and participants with abnormal colposcopy received histopathology. During the study, participants with histopathological findings ≥CIN2 discontinued follow-up. as detailed in Before program implementation, standard training was provided for cervical exfoliation cytology sampling (for cytology testing and HPV testing), stain production, reading, laboratory testing, colposcopy, and histopathology. All program staff were required to pass an assessment by the designated medical institution. Senior physicians from the Heping Hospital of Changzhi Medical College were on site for long periods of time to provide quality control guidance during the program.in vitro amplification and DNA reverse spot hybridization method to detect 14  high-risk human papillomavirus nucleic acid subtypes.HPV typing nucleic acid detection kit (biochip method)  uses a nucleic acid amplification-reverse spot hybridization technique combining PCR The HPV DNA typing test was performed using the high-risk HPV5 + 9 nucleic acid test kit (Tellgen Corporation). The test kit is based on fluorescent PCR technology and detects HPV 16, 18, 33, 52, 58, and 9 other HPV  typing for a total of 14 HPV and human genomic β-Globin DNA in a single reaction tube using a four-channel fluorescent PCR instrument with a minimum detection limit of 100 copies/ test, simultaneous detection of cervical exfoliated cell gene β-Globin.All of the following tests and diagnostic procedures were strictly double-blinded. Cytology slides were read by two pathologists results and reported according to the Bethesda 2014 classification. The cytological results were as follows: negative for intraepithelial lesion or malignancy (NILM), atypical cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells–cannot exclude high-grade squamous intraepithelial lesion (ASC-H), high-grade squamous intraepithelial lesion (HSIL), atypical glandular cells (AGC), and cervical cancer cells. Diagnoses were reported if the diagnoses by two cytologists were consistent. Otherwise, a third cytologist was consulted.All participants with positive HPV 18 and/or HPV 16 testing or abnormal cytology (ASC-US or worse) were referred to undertake colposcopy. Two pathologists independently made diagnoses according to the 2014 WHO Classification of Tumors of the Female Genital Tract. If the diagnoses were concordant, they were reported as the pathologic diagnosis. Otherwise, a panel of pathologists was consulted to reach a consensual diagnosis. The histological diagnoses of cervical lesions were classified as normal, LSIL, HSIL/CIN2, HSIL/CIN3. HPV testing, cytology, and pathologic examination were performed with blinding to the results of each test. Participants with both negative HPV and cytology results were not referred for colposcopy and were considered normal.p < 0.05(two-sided) was considered statistically significant. Using cervical intraepithelial neoplasia grade 2 or worse as a reference standard, sensitivity, specificity, positive predictive value, negative predictive value, and area under the receiver operating characteristic (ROC) curve (AUC) were calculated.In this study, an ACCESS database was developed, double-entered and checked until complete consistency before analysis. Data were analyzed using R software (version 4.1.2). We reported numbers and percentages for categorical variables. Chi-square test were used to compare the difference between groups. Based on a review of the published literature , we chosA total of 3,328 participants were included in the HPV PCR testing cohort, with an HPV prevalence of 14.90%. The majority of the participants were aged 40 years or above. About half of them completed secondary education, and almost all of them were married, non-smokers, and non-drinkers. There were no statistically significant differences in different subgroups of age at menarche, age at first pregnancy, number of pregnancies, and number of births.A total of 3,919 participants were included in the HPV DNA chip testing cohort, with an HPV prevalence of 21.10%, and the distribution of population characteristics was similar to that of the HPV testing cohort and was overall comparable, as shown in χ2 = 186.85, p < 0.001). In the HPV DNA chip population, a total of 26 cases of CIN2+ were identified at baseline, all of which were found in the HPV-positive population . Among the 4-year cumulative cases, a total of 53 cases of CIN2+ were found in the population with HPV PCR testing, 51 of which were in the HPV-positive population . Fifty-four cases were found cumulatively in the HPV DNA chip population, all of which were in the HPV-positive population , and all of these differences were statistically significant.CI: 64.27, 94.95) and 77.36 , respectively. There was a relatively good improvement in sensitivity and specificity when LBC was combined with HPV testing for screening. The sensitivity of both strategies 3 and 4 decreased when using CIN2+ as the gold standard. However, precancerous lesions are a progressive state. Therefore, there was a significant improvement in sensitivity and specificity when using the 4-year cumulative CIN2+ as the gold standard. For strategies 3, 4, 5 and 6, HPV DNA chip technology performed better than PCR HPV test. The same trend was shown for CIN2+ over the next 4 years  CIN2+ cases were found cumulatively over 4 years, while in the chip cohort, 54 (1.38%) cases were found cumulatively over 4 years, with no statistically significant differences. In the present study, DNA microarray outperformed PCR performance. The DNA microarray performed better when LBC was combined with HPV screening, both for baseline CIN2+ and cumulative CIN2+ over 4 years. Differences in detection principles may be an important reason for this difference. The sensitivity of LBC (ASC-US+) in the two cohorts in this study was 55.26 and 84.62%, respectively. The difference in cytology sensitivity between the PCR and chip cohorts may be related to the performance of cytology physicians and field sampling, and it is important to note that these physicians were from low to moderate resource areas.In a previous study that pooled 40 studies, it showed that different principles of HPV testing perform differently in screening . HC2 is The advantage of this study is the ability to see the long-term effects of screening through 4-year follow-up. Cytology sampling and diagnosis are from rural areas, which more realistically reflects the performance of cytology in areas with poor and moderate resources, but this is also a disadvantage. In addition, the cytology performance of the two cohorts was inconsistent, which may be related to the inconsistent performance of physicians in the regions or because of differences in population characteristics. Thirdly, as colposcopy was not performed in our programme in the cytologically normal and HPV-negative population, this may have led to partial under-diagnosis. Furthermore, colposcopic images were not reviewed and the subjectivity of the colposcopist can influence sampling. Finally, health resources and demographic characteristics vary widely across different provinces in China. Also, the cohort population was from two counties in Shanxi province, which is not fully representative of rural China. Therefore, the results of this study need to be further validated in other rural areas.More studies should be conducted in the future to confirm the performance of HPV DNA chip. The development of HPV DNA chip with low price and high performance could provide more options for HPV test-based cervical cancer screening.In conclusion, we selected two rural areas in China for a four-year follow-up to evaluate cytology and different HPV testing principles and concluded that HPV testing is more effective than LBC for screening and HPV DNA microarray testing is more effective than HPV PCR testing. Optimal screening can be achieved when used in combination with LBC for ASC-US population triage, if consideration is given to the feasibility of resource availability.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by Affiliated Heping Hospital of Changzhi Meidical College. The patients/participants provided their written informed consent to participate in this study.Y-LQ and X-XF contributed to the design. Z-FL and X-HJ wrote the manuscript. B-KW, X-YR, and WC performed statistical analysis and HPV test. L-BW contributed to cytology and histology examination. All authors read and approved the final manuscript.This research was supported by CAMS Innovation Fund for Medical Sciences (no. CAMS 2021-I2M-1-004).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Perennial grain crops could make a valuable addition to sustainable agriculture, potentially even as an alternative to their annual counterparts. The ability of perennials to grow year after year significantly reduces the number of agricultural inputs required, in terms of both planting and weed control, while reduced tillage improves soil health and on-farm biodiversity. Presently, perennial grain crops are not grown at large scale, mainly due to their early stages of domestication and current low yields. Narrowing the yield gap between perennial and annual grain crops will depend on characterizing differences in their life cycles, resource allocation, and reproductive strategies and understanding the trade-offs between annualism, perennialism, and yield. The genetic and biochemical pathways controlling plant growth, physiology, and senescence should be analyzed in perennial crop plants. This information could then be used to facilitate tailored genetic improvement of selected perennial grain crops to improve agronomic traits and enhance yield, while maintaining the benefits associated with perennialism. Climate change has led to an increased focus on sustainable agricultural practices to produce food and feed using less energy and with a lower carbon footprint , rice (Oryza sativa), and wheat (Triticum aestivum) account for 28% of global primary crop production  and banana (Musa sp.), which can produce 50 t ha−1 of starchy fruit per year , and sorghum (Sorghum bicolor), delays in senescence or “staygreen” traits are associated with stress tolerance , staygreen traits are associated with reduced tillering, deeper roots, altered stomatal conductance, and improved nitrogen dynamics  plants were stored as stem fructan and not remobilized. Similarly, a study in maize (Z. mays) found that while post-anthesis nitrogen uptake of staygreen lines was greater compared to non-staygreen types, nitrogen remobilization to the grain was lower  plants were significantly greater when compared to 2-year-old plants   is widespread and generally more prevalent in perennial crops compared to annuals , the S locus was mapped to homoeologous group 1 and was found to encode a domain of unknown function (DUF) gene , the Z locus was mapped to homoeologous group 2 and shown to encode another domain of unknown function gene, DUF247, alongside a closely linked ubiquitin-specific protease (USP) , similar SI mechanisms appear to be involved, whereby assessment of progeny combinations resulting from a polycross led to the identification of S and Z loci gene candidates  to great success  with perennial S. strictum resulted in multivalent formation during meiosis and chromosomal translocations, reducing fertility  to autoallodecaploid (2n = 10x = 70), with aneuploid and amphidiploid events routinely reported  mapping ; Table 1O. sativa and perennial O. longistaminata species have led to the identification of multiple QTLs relating to rhizome formation  x S. strictum  population identified five QTLs accounting for 74% of variation in perennation , such a narrow definition of the perennial trait may not account for wider physiological differences associated with the differences in life cycles. For example, genetic analysis conducted using populations derived from crossing annual Z. mays with its perennial relative Z. diploperennis led to identification of QTLs associated with tillering  , has been identified as the primary regulator in the perennial species Arabis alpina  analysis has been conducted between the two species to identify genetic differences   and the rhizomatous perennial relative, O. longistaminata  with perennial S. halepense , a perennial grass that has mostly been utilized for forage but is now being developed for use as a perennial grain crop through de novo domestication. Currently, the crop is produced at a small scale in the USA, sold under the trade name Kernza®, and incorporated into specialty products   have developed rapidly in the last few years, utilizing a wide range of ground-based sensors or airborne platforms, including drones, for the collection of phenotypic and climatic data  species share commonalities     and BABY BOOM (BBM) have been shown to improve the regeneration frequency of a variety of transformation recalcitrant annual grain crops, including maize, sorghum, and indica rice (O. sativa ssp. indica)  results in increased levels of GRF4 transcripts, contributing to the enlarged grain size of transgenic rice lines  could also be used to modulate the expression of both homologous and homoeologous gene copies. MicroRNAs are small molecules that target specific mRNA targets based on their sequence complementarity. Through miRNA binding, mRNAs are targeted for degradation or repression of translation, contributing to the regulation of developmental and physiological processes. In rice, the disruption of miR396 recognition sites in H. bulbosum will be briefly outlined, while those related to T. intermedium (Kernza®) have been reported elsewhere . Accessions evaluated were primarily sourced through the Nordic Genetic Resource Centre (NordGen) and selected to represent the species' original distribution from regions around the Mediterranean Sea to Central Asia. The best performing genotypes have been selected, with offspring used for breeding and development of perennial barley.For the major crop species H. bulbosum, the elimination of H. bulbosum chromosomes is common, resulting in barley haploids , and is outcrossing  genomes, containing highly repetitive and complex regions that make whole-genome sequencing and assembly challenging. However, in 2017, the International Barley Genome Sequencing Consortium (IBSC) used hierarchical shotgun sequencing to successfully assemble and sequence the genome of H. vulgare cv. Morex , the Innovation Fund Denmark , and the Novo Nordisk Foundation .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Humility and modesty are both emphasized in Eastern and Western societies. However, people usually conflate them in everyday usage. To reduce the confusion of the two constructs, it is very vital to carefully differentiate the two constructs and examine whether they lead to similar or different effects on job performance. In this study, we scrutinized the effects of the two constructs on four dimensions of job performance simultaneously, including task performance, citizenship behavior (helping and voicing), unethical pro-organizational behavior, and innovative behavior. Using a dataset of 239 employees and 77 supervisors, we showed that modesty is not related to task performance and voicing, but that it is positively related to unethical pro-organizational behavior and negatively related to helping and innovative behavior. In contrast, we showed that humility is negatively related to unethical pro-organizational behavior and positively related to task performance, helping, voicing, and innovative behavior. Our findings reveal that modesty and humility can lead to very divergent work outcomes. The results strongly support the idea that modesty and humility are distinct constructs embedded in separate nomological networks and strongly suggest that organizations should encourage employees’ humility rather than modesty. The theoretical and practical implications of this work are discussed. Individual work role performance drives the success of organizations and even the entire economy . Not surAs important positive character strengths and virtues, both humility and modesty have received increasing attention in organizational research recently because they are theoretically expected to bring organizations positive outcomes . Indeed,However, humility and modesty are quite similar . Lay peoDo humility and modesty lead to similar or different work outcomes? To answer this question, we first reviewed and differentiated the two constructs. To more clearly and cleanly differentiate the two constructs, we define the two constructs from the interpersonal perspective because the overlap between modesty and humility mainly lies in the intrapersonal aspect. In the present study, humility (qianxu in Chinese) refers to interpersonal behavioral manifestations that reflects a person’s willingness to acknowledge personal limitations or mistakes, appreciate others’ strengths, and be open to feedback and advice from others  and violate core societal values, mores, laws, or standards of proper conduct” , p. 622.In this study, we posit that humility and modesty may affect employees’ UPOB differently. According to the theory of moral self, individuals with strong moral identities are particularly sensitive and reactive to ethical issues compared with those with weak moral identities . Indeed,Instead, modest behaviors are mainly driven by a person’s desire to protect themselves , rather Hypothesis 4: Humility is negatively related to unethical pro-organizational behavior, whereas modesty is positively related to unethical pro-organizational behavior.Innovation has been especially emphasized in this knowledge-based era because of its central role in the long-term survival of organizations. Innovative behavior refers to “the intentional proposal and application of novel and improved ideas, processes, practices, and policies aimed at organizational effectiveness, business success, and long-term sustainability” , p. 2. WSecond, humility can boost innovation by making people more empathetic. Empathy, which refers to the ability to share the feelings of another, is especially necessary for customer-centric innovation . HumilitIn contrast, we posit that modesty will negatively correlate with innovative behavior. First, innovation means doing things differently and challenging the status quo , which wHypothesis 5: Humility is positively related to innovative behavior, whereas modesty is negatively related to innovative behavior.We first recruited 77 managers who opted to voluntarily participate in our survey. All the managers were former or current MBA students at an Eastern China university. We then asked them to randomly select a maximum of five subordinates who would like to voluntarily participate in our survey. After obtaining consent from the subordinates, the managers provided us with their subordinates’ names. We recruited 261 subordinates. We then created the questionnaire code for each subordinate and manager, which was used to match the manager–subordinate questionnaires. Each questionnaire was sent to the participants in an envelope and with a note reminding them to put the completed questionnaire in the envelope and seal it before returning. In addition, in every questionnaire, we clearly stated the confidentiality of the academic survey and that all the data would be calculated in aggregation. We also stated clearly the voluntary nature of the survey and reminded the participants that they could withdraw at any time.To test our hypotheses, we collected job performance data from both the managers and the subordinates. The survey was conducted in two waves. In the first wave, we sent the managers a questionnaire to ask them to rate their subordinates’ humility and modesty. One month later, we sent the managers the second-wave questionnaire, in which we asked them to rate their subordinates’ task performance, helping, voicing, UPOB, and innovative behavior. Meanwhile, we also sent the subordinates a questionnaire, in which we asked them to rate their own task performance, helping, voicing, UPOB, and innovative behavior. Finally, we collected 239 manager–subordinate dyadic datasets. The final employee sample consisted of 62.8% men and 37.2% women. Eighty-nine percent of the subordinates had a college degree or above. The mean organizational tenure of the subordinates was 3.47 years (SD = 2.89). The proportion of subordinates aged <26 years, 26–45 years, and ≥ 46 years was 17.6%, 59%, and 23.4%, respectively. With regard to industries, about 36.4% participants worked in IT and software service industry, 28% worked in manufacturing industry, 19.2% worked in banking and financial service industry, 9.2% worked in advertising industry, and 7% worked in trading and other industries.The managers reported their subordinates’ humility, modesty, task performance, helping, voicing, UPOB, and innovative behavior. In addition, the subordinates reported their own task performance, helping, voicing, UPOB, and innovative behavior. All the questionnaires were administrated in Chinese. All the original English questionnaires were translated into Chinese using a standard back-translation procedure .Humility was assessed using the nine-item scale proposed by We measured modesty using the 3-item scale from Task performance was assessed using the six-item measure from Helping was assessed using the 7-item measure by Voicing was assessed using the 10-item measure from UPOB was assessed using the six-item measure proposed by Innovation is usually defined as a process involving both the generation and implementation of ideas. Thus, we adopted the 6-item measure by Studies have shown that there are gender and age differences in modest self-presentation . Hence, To test our hypothesized model, we used Mplus 8.0 to conduct the statistical analysis. First, we conducted a series of confirmatory factor analyses (CFA) to confirm construct validity. Next, we conducted structural equational modeling (SEM) to test our hypotheses. As the supervisors were required to rate several subordinates’ job outcomes, we controlled for the potential nesting effect in all the CFA and SEM analyses.X2(48) = 105.63, p < 0.01, comparative fit index (CFI) = 0.94, root-mean-square error of approximation (RMSEA) = 0.07, standardized root-mean-square residual (SRMR) = 0.05. All the factor loadings ranged from 0.69 to 0.91. In addition, the three dimensions of humility were significantly correlated . However, modesty was only related to the first dimension of humility , but it was not related to appreciation of others’ strengths  or openness to feedback .To identify whether modesty is a distinct construct from humility, we first constrained the items for modesty and three dimensions of humility to load on their respective factors. We found that the four-factor model had an adequate fit: X2(50) = 120.05, p < 0.01, CFI = 0.93, RMSEA = 0.08, SRMR = 0.07], it was significantly worse than the four-factor model . In addition, the results showed that the general factor was strongly related to admitting personal limitations , appreciation of others’ strengths , and openness to feedback , but it was not related to modesty , suggesting that modesty and humility were different constructs.We then tested a second-order model with a general factor to predict modesty and three dimensions of humility. X2(461) = 693.83, p < 0.01, CFI = 0.95, RMSEA = 0.05, SRMR = 0.05], much better than any other alternative model, including the one-factor model . Finally, we conducted a series of CFAs on the 12 studied variables to examine their discriminative validity. Too many items may decrease the sample-size-to-parameter ratio; however, parceling can help to alleviate this problem . We randTo alleviate common method bias, we collected data from multi-sources and in multi-waves. However, since supervisors reported both independent and dependent variables, we still conducted Harman’s single-factor test to evaluate the common method variance. Specifically, we conducted exploratory factor analysis (EFA) for all items from the supervisor-rated constructs. EFA indicated that 32.79% (less than 50% of the recommended threshold) of the total variation was extracted by one factor. Thus, there is no problem with common method bias in our study.r = 0.14, p < 0.05; supervisor-rated: r = 0.28, p < 0.01) and negatively related to innovative behavior . Humility was positively related to performance , helping , voicing , and innovative behavior , but it was negatively related to UPOB . These initial findings supported Hypotheses 4 and 5 and partial support for Hypotheses 1–3.X2 = 887.13, p < 0.01, degrees of freedom [df] = 557, RMSEA = 0.05, CFI = 0.94, SRMR = 0.05). The results for the proposed paths are exhibited in To test our proposed hypotheses, we conducted SEM analyses using Mplus version 8.0. Our proposed model fitted the data very well . However, modesty was not significantly related to task performance, although the coefficients were negative. Hence, Hypothesis 1 was only partially supported.Hypothesis 1 states that humility is positively related to task performance, whereas modesty is negatively related to task performance. Our results revealed that humility was positively related to task performance . Despite modesty was not significantly related to supervisor-rated helping, it was negatively related to employee self-reported helping . Hence, Hypothesis 2 was marginally supported.Hypothesis 2 states that humility is positively related to helping, whereas modesty is negatively related to helping. Our results showed that humility was positively related to helping . However, modesty was not significantly related to voicing, although the coefficients were negative. Hence, Hypothesis 3 was only partially supported.Hypothesis 3 states that humility is positively related to voicing, whereas modesty is negatively related to voicing. Our results showed that humility was positively related to voicing , although it was not related to employee self-rated UPOB. In addition, modesty was positively related to UPOB . Hence, Hypothesis 4 was marginally supported.Hypothesis 4 states that humility is negatively related to UPOB, whereas modesty is positively related to UPOB. Our results showed that humility was negatively related to supervisor-rated UPOB . In addition, humility was positively related to innovative behavior . Thus, Hypothesis 5 was supported.Hypothesis 5 states that humility is positively related to innovative behavior, whereas modesty is negatively related to innovative behavior. Our results showed that modesty was negatively related to innovative behavior  that is salient in the Chinese context but has been neglected in the previous studies. Studies on innovation have identified various barriers . In thisFinally, we contribute to the UPOB literature by revealing that modesty can foster UPOB, but humility can inhibit it. Studies on the antecedents of UPOB have mainly paid attention to situational factors, such as leadership styles, and have seldom focused on individual factors . In addiThis study has several important managerial implications. First, our findings revealed that employees’ humility and modesty have divergent effects on job performance. More specifically, humility had a positive effect on task performance, citizenship behavior, and innovative behavior, and a negative effect on UPOB. In contrast, modesty had a negative effect on helping and innovative behavior and a positive effect on UPOB. Given that people usually conflate humility and modesty, our research shows that managers should differentiate between them because they have very different predictive effects.Second, Chinese culture especially emphasizes modesty . StudiesThis study has some limitations that should be noted. First, this study was conducted in the context of Chinese culture. In Western and Eastern cultures, people may have different norms and attitudes toward modesty and humility ; thus, oSecond, we only investigated the direct effects of modesty and humility on job performance, and we did not directly investigate the underlying mechanism or the boundary conditions. We believe that there are various individual and situational factors that may influence the effects of modesty and humility on work outcomes. For example, future research could explore the potential moderating effects of political skills and different leadership styles.Third, we could not rule out the effects of previous job performance as we did not measure the previous job performance in time 1. Future research could include more control variables to rule out other explanations. In addition, our design was still correlational despite we measured independent variables and dependent variable at two times. Future research could employ experiment method to examine our research model.Using multi-wave and multi-source data from China, we simultaneously investigated the effects of employees’ humility and modesty on job performance. Our results suggest that modesty is not a sub-domain of humility. Compared with humility, modesty has different effects on work outcomes. Our results reveal the importance of differentiating between humility and modesty. They also suggest that organizations should encourage employees’ humility rather than modesty. Our study emphasizes that organizations, especially Chinese organizations, should not have a naïve notion of employees’ modesty.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by the Research Ethics Committee of Department of Psychology, Fudan University. The patients/participants provided their written informed consent to participate in this study.HP conceptualized the study, collected the data, analyzed the data, drafted the manuscript, and approved the submitted version.This research was funded by the National Natural Science Foundation of China (grant no. 71772047).The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Overall survival for HPV-associated oropharyngeal squamous cell carcinoma (OPSCC) has differed by sex, but little is known regarding cancer-specific outcomes. We assessed the independent association of sex with cancer-specific survival in patients with HPV-associated oropharyngeal squamous cell carcinoma (OPSCC).  We identified 14,183 patients from the Surveillance, Epidemiology, and End Results (SEER) program with OPSCC and tumor HPV status. We used Kaplan–Meier methods to compare overall survival (OS) and OPSCC-specific survival (HNCSS) by patient sex and by tumor HPV status. We then separately fit multivariable survival and competing risk models evaluating the association of sex on these outcomes by tumor HPV status and stratified by the use of guideline-concordant OPSCC treatment. p < 0.001). Women with HPV-positive tumors were less likely to receive guideline-concordant treatment compared to men. In unadjusted survival analyses, women did not differ in OS or HNCSS compared to men for HPV-positive tumors but had worse OS and HNCSS for HPV-negative tumors. After adjustment, men and women with HPV-positive OPSCC did not differ in OS or HNCSS. However, women with HPV-negative tumors faced worse overall survival (hazard ratio (HR) 1.15, 95% CI 1.02–1.29) that persisted even after stratifying for stage-appropriate treatment .  A total of 10,210 persons with HPV-positive tumors (72.0%) and 3,973 with HPV-negative tumors (28.0%) were identified. A larger proportion of women had HPV-negative tumors (24.0%) versus HPV-positive tumors (13.2%;  Women with HPV-positive OPSCC had similar survival outcomes compared to men, but those with HPV-negative tumors have worse overall and cancer-specific survival. For the past 4 decades, the incidence of oropharyngeal squamous cell carcinoma (OPSCC) has steadily increased, growing by 122% in men and 43% in women from 1999 to 2015.  OPSCC isIncidence and temporal trends related to HPV-associated OPSCC specifically have also differed. Recent estimates have shown that HPV-associated OPSCC has increased annually 0.8% for women and 2.7% for men.  AlthoughOPSCC p16 status has been strongly associated with treatment responsiveness and survival outcomes. –16 HowevPrevious OPSCC outcome studies have been limited by sample size or lack of data on cause of death, limiting the analysis to overall survival only, and, therefore, are susceptible to bias related to noncancer causes of death related to comorbid illnesses and behavioral factors. In addition, large cancer registry studies have lacked information on HPV status for OPSCC cases. In this study, we used data from a contemporary cohort of OPSCC cases from a national representative cancer registry database to compare overall and cancer-specific survival by sex, according to tumor HPV status. Our hypothesis is that women with OPSCC will have better survival outcomes regardless of HPV status, in line with trends seen in other cancers overall.We performed a retrospective cohort study using data from the Surveillance, Epidemiology, and End Results (SEER) Head and Neck with HPV Status Database. The SEER program has collected clinicopathologic data on incident cancer cases from 18 population-based registries, which represent approximately 28% of the US population. The Head and Neck with HPV Status Database includes cases diagnosed between 2010 and 2016, for which available HPV data have been collected.  Tumor HPInternational Classification of Diseases for Oncology, Third Edition (ICD-O-3), topography codes C01.9–10.9, which includes the base of the tongue, lingual tonsil, soft palate, uvula, tonsillar fossa, tonsillar pillar, tonsil, vallecula, anterior surface of the epiglottis, lateral and posterior wall of the oropharynx, branchial cleft, and oropharynx, not otherwise specified. Cases that were not of squamous cell histology, as determined by ICD-O-3 (8050–8084), were excluded. Next, we identified and included cases with known American Joint Committee on Cancer, 7th edition, clinical stage, which we restaged to the 8th edition based on available T, N, and M information on the individual level. Cases with unknown HPV status were excluded . A total of 14,183 patients were identified  or HPV negative .We identified incident cases of OPSCC using the entified . We thenT, N, and M values) and treatment type were ascertained from SEER on the individual level. We defined surgery as any definitive resection surgery performed, including excisional surgery of the oropharynx, the base of the tongue resection, palatectomy, uvulectomy, and/or tonsillectomy. Radiotherapy was coded for cases receiving beam radiation or a combination of a beam with implants or isotopes. Chemotherapy was included in the database and encoded as “yes” or “no/unknown.” Each case was then encoded with all treatments received, whether surgery, radiotherapy, or chemotherapy alone or in combination with other treatment methods. To determine whether the guideline-concordant treatment was administered by clinical stage, treatments were compared to National ComprehensiveCancer Network (NCCN) guidelines. [The SEER data were used to identify age, race/ethnicity, year of OPSCC diagnosis, median household income by county attribute, proportional educational attainment by county attribute, and insurance type . Tumor characteristics , cancer stage, and treatment by tumor HPV status and stratified by the use of guideline-concordant OPSCC treatment. We separately compared outcomes for all HPV-positive and HPV-negative OPSCC cases, then, stratified by tumor stage, and then, further stratified by use of guideline-concordant treatment, using the aforementioned models. We also tested for interactions between sex and tumor stage. Last, to evaluate the effects of potential differences in competing (non-OPSCC-related) causes of death by sex, we fit similar multivariable Fine-Gray regression models, stratified by HPV tumor status, first for all cases, and then limited to patients receiving guideline-concordant treatment. Complete models are provided in Supplementary Tables. All analyses were performed in STATA Version 13 . STATA code for all analyses can be found online at this link: https://github.com/derkao/opscc-by-sex. The Mount Sinai Institutional Review Board determined that this study was exempt from human research.We tested for differences in baseline and clinical characteristics between women and men stratified by tumor HPV status using the p < 0.001). For HPV-positive OPSCCs, in comparison to men, women were older (p < 0.001), more likely to be non-Caucasian (p=0.001), early stage (p < 0.001), single (p < 0.001), and more likely to be insured with Medicaid (p < 0.001). Among HPV-negative OPSCC cases, women were also older compared to men at diagnosis (p < 0.001), earlier stage (p < 0.001), and more likely to be single (p < 0.001).In total, 14,183 cases of OPSCC with known staging and HPV status were identified, of which 10,210 (72.0%) were HPV positive and 3,973 (28.0%) were HPV negative . Women wp=0.002) compared to men. For HPV-negative tumors, a disparity of similar magnitude was seen; however, the comparison was not significant (p=0.06). Differences in the use of guideline-concordant treatment for women with HPV-positive tumors appear to stem from less aggressive treatment for earlier-stage (I-III) cancers. In contrast, for HPV-negative tumors, women were less likely to receive guideline-concordant treatment for more advanced-stage cancers . After adjusting for sociodemographic factors, including race, median county household income, proportional county education attainment, insurance type, cancer stage, and marital status, sex was still a significant predictor of guideline-concordant treatment for HPV-positive tumors  but not HPV-negative tumors (p=0.09).In univariate analyses, women with HPV-positive OPSCC were significantly less likely to receive guideline-concordant treatment ; p=0.002p = 0.02).In unadjusted survival analyses , men and women with HPV-positive tumors did not differ in overall survival (OS)  or head and neck cancer-specific survival (HNCSS) , defined by death due to cancer in any head and neck structure. However, in HPV-negative cases, women had worse OS  and HNCSS . Worse cancer-specific survival persisted for women after accounting for possible competing risks . Among patients who received guideline-concordant treatment, we found similar trends; women had no difference in outcomes for HPV-positive tumors but had significantly worse HNCSS for HPV-negative tumors even after accounting for competing risks  compared to head and neck cancer treatment by sex and found that women received less intensive treatment, despite worse cancer-specific survival. [National Cancer Database, patterns of treatment for HPV-positive and HPV-negative OPSCC differed for women versus men; however, proportions of untreated patients were similar. [We found differences in treatment patterns for women and men with OPSCC. For both HPV-positive and HPV-negative tumors, women were less likely to receive guideline-concordant treatment. A recent large cohort study  or environmental exposures, which may have impacted cancer outcomes. Fourth, missing HPV data may be a source of selection bias, as previous studies have demonstrated disparities of HPV testing by demographics such as race and insurance.  Lastly, In this study of population-based cancer data, we found that outcomes for OPSCC did not differ for women compared to men for HPV-positive tumors. Women, however, did experience worse overall and cancer-specific survival for HPV-negative tumors, which was not fully explained by treatment disparities.


Random compound screening identified small molecules with Emax values similar to 8-bromoguanine. Paraquat-induced mitochondrial dysfunction was attenuated by several small molecule OGG1 activators; benefits included enhanced mitochondrial membrane and DNA integrity, less cytochrome c translocation, ATP preservation, and mitochondrial membrane dynamics. Our results support an allosteric mechanism of OGG1 and not product-assisted catalysis. OGG1 small molecule activators may improve mitochondrial function in oxidative stress-related diseases.8-Oxoguanine DNA glycosylase (OGG1) initiates base excision repair of the oxidative DNA damage product 8-oxoguanine. OGG1 is bifunctional; catalyzing glycosyl bond cleavage, followed by phosphodiester backbone incision via a β-elimination apurinic lyase reaction. The product from the glycosylase reaction, 8-oxoguanine, and its analogues, 8-bromoguanine and 8-aminoguanine, trigger the rate-limiting AP lyase reaction. The precise activation mechanism remains unclear. The product-assisted catalysis hypothesis suggests that 8-oxoguanine and analogues bind at the product recognition (PR) pocket to enhance strand cleavage as catalytic bases. Alternatively, they may allosterically activate OGG1 by binding outside of the PR pocket to induce an active-site conformational change to accelerate apurinic lyase. Herein, steady-state kinetic analyses demonstrated random binding of substrate and activator. 9-Deazaguanine, which can’t function as a substrate-competent base, activated OGG1, albeit with a lower E Among various oxidative DNA products, 8-oxoguanine  with several age-related diseases including cancer, neurodegeneration, chronic obstructive pulmonary disease, diabetes, and heart failure13.Genomic DNA, particularly mitochondrial (mt), is prone to oxidative attack and modification by reactive oxygen species (ROS). Endogenous ROS arise primarily as by-products of mitochondrial oxidative phosphorylation or from exogenous exposure to chemicals and physical agents such as ionizing radiation16. 8-Oxoguanine DNA glycosylase (OGG1) is a bifunctional enzyme that catalyzes both a glycosylase and an AP lyase reaction. OGG1 performs the first, and perhaps also the rate-limiting step, in the damage control pathway involving 8-oxo-dG17. Several studies have demonstrated that the overexpression of OGG1, particularly within the mitochondrial compartment, confers cellular protection in assorted cellular and animal models; for examples, see21. These findings support the view that accelerating OGG1 activity, especially within mitochondria, may be beneficial to mitochondrial function by minimizing the accumulation of mutagenic and cytotoxic 8-oxo-dG in DNA during periods of oxidative stress.To counteract these potentially mutagenic consequences, cells primarily use base excision repair (BER), which is a complex multi-enzymatic process that is accomplished through the coordinated action of a DNA glycosylase, AP endonuclease, polymerase-β (or polymerase-γ in the mitochondria), as well as a DNA ligase22. This attack leads to the formation of a Schiff base adduct between the ε-amino group of Lys249 and C1′ of the abasic product  product is retained at the product recognition pocket. The N9-atom of the glycosylase product then serves as the catalytic base for C2′-proton abstraction, which is the trigger step for the AP lyase reaction  and 8-aminoguanine  binding of 8-bromoGua in the catalytic pocket would competitively inhibit enzyme activity; 2) product-assisted catalysis may be interesting mechanistically but may not be relevant in vivo and 3) 8-bromoGua might be an allosteric activator of AP lyase activity although structural support for the latter was lacking. However, binding of free 8-oxoGua at a non-substrate site on OGG1 has been shown to occur with sub-nM affinity and produced a conformational change as measured by intrinsic OGG1 tryptophan fluorescence26. Thus, binding of an activator with free enzyme at the catalytic site would prevent binding of substrate and would result in enzyme inhibition rather than activation. On the other hand, direct binding to free enzyme is consistent with allosteric activation in which 8-oxoGua or its analogues bind to a site outside of the nucleobase recognition pocket to induce a conformational change in the active site. This would move an otherwise misplaced, enzyme-derived base group into a proximal position for C2′-proton abstraction  with a fluorescent quench pair of 5′-FAM and 3′-DAB , or by a combination of the two (α < 1 and β > 1). For an ordered mechanism with substrate binding first . Alternately, in an ordered mechanism with activator bound first , the β value, which measures the fold of kcat activation, is 18 ± 3 which is 50% of the catalytic activity of OGG1 in the presence of 8-bromoGua  were similar to the Emax value obtained for 8-bromoGua, which served as the positive control. These data indicated that the activation efficiency of these compounds, which were structurally distinct from product analogs, was as great or greater than that of 8-bromoGua. It is difficult to reconcile this outcome by product-assisted catalysis. Rather the results can be easily explained by allosteric activation, in which activation is not achieved through direct involvement of activator in catalysis, but by a conformational change at the active site upon binding of activator to an allosteric binding pocket. The latter step reorients the enzyme-derived catalytic base group for C2′-proton abstraction , the 19F NMR resonance was broadened while also being shifted up field. This finding demonstrates that this compound binds to the OGG1 protein in the absence of a DNA substrate. Figure VII in the presence of compound VI and OGG1. The line narrowing observed on the 19F NMR resonance of compound VI suggests that compound VII competes for the same binding site as compound VI. Similar results were obtained with compounds VIII-X as shown in Fig. 19F NMR resonance of compound VI. These results further support an allosteric activation mechanism.Compound I up to 2 μM, the I up to 2 μM, the Mixed probe molecular dynamics studies identified multiple sites which could accommodate small molecule ligands with appreciable affinities (< 10 μM). Docking studies were performed against seven of the largest sites  exposure, total glutathione decreased from 21.59 ± 0.44 mM to 17.25 ± 1.14 mM or by 20.1% but after 48 h, it declined to 3.9 ± 0.07 mM or by 87.5%. Total cellular glutathione loss indicated the severity of the ROS production throughout the cells even though it seemingly emanated from within the mitochondrial compartment. The ROS generated in the mitochondria also impacted nuclear DNA by inducing nuclear double-stranded breaks that were detected by γ-H2-AX staining. These breaks were observed only at 48 h beginning at 0.1 mM PQ  provoked a concentration-dependent rise in this important parameter  and VII (30 μM) essentially blocked this PQ (0.6 mM)-induced change. The discrepancy between the efficacy of compound XIII noted in the latter experiment and the lack of efficacy with mtDNA damage is not readily apparent at this time. Compound XIII was effective in preventing accumulation of 8-oxo-dG under similar conditions27.Although it can’t be ruled out that PQ stimulated ROS in areas other than the mitochondria, the physical tethering of the mtDNA to the inner mitochondrial membrane, which houses Complex I, renders both the mitochondrial membranes and the mtDNA a likely ROS target. The consequences of excessive ROS production on mitochondrial membrane integrity are depicted in Fig. ter Fig. d. Maximae c Fig. h,i. Pane2O2 to lower mtDNA content  indicated that the fall in ATP and the rise in ADP provoked by PQ was partially attenuated. The levels of both energy metabolites were increased across the PQ concentration range in the presence of the activators. For example, at 3 mM PQ, the ATP concentration was 0.63 ± 0.05 μmol/gm protein with PQ alone but was 1.31 ± 0.11 μmol/gm protein for PQ plus compound XI. Similar findings were obtained for compounds XIII and VII, respectively, i.e., 0.96 ± 0.10 vs 1.37 ± 0.30 and 0.61 ± 0.09 vs 1.73 ± 0.08 μmol/gm protein. These differences at this PQ concentration represented increases of 108%, 43% and 184%, respectively, which were marked improvements in the amount of ATP. The differences in [ADP] between PQ (3 mM) alone and PQ plus activators were only 9, 13 and 54%, respectively. By diminishing the impact of PQ on mitochondrial bioenergetics, the OGG1 activators maintain cell health, consistent with our earlier results27.Mitochondrial bioenergetics in A549 cells in response to PQ produced time- and concentration-dependent changes Table . At 24 h35. The translocase of the outer mitochondrial membrane  is responsible for the import of nuclear-encoded proteins into the mitochondria. In the present study, PQ (0.03–3 mM) resulted in varied changes in TOM20 immunofluorescence (overlaid with images of MitoTracker™ orange and Hoechst 33342) in A549 cells. At low PQ concentrations (0.03–0.3 mM) TOM20 immunofluorescence increased to nearly 25% above baseline after 48 h of incubation of PQ at 0.3 mM PQ but then abruptly dropped below baseline values at 0.6–3 mM PQ  resulted in markedly higher values. PQ plus compound XII showed a concentration-dependent rise in TOM20 immunofluorescence whereas compound XIII  and VII (25 μM) had few effects. The impact of PQ plus or minus the OGG1 activators on DRP1 content using quantitative imaging is shown in Fig. XI, XII, XIII and VII, applied 4 h prior to PQ with 0.6 mM PQ, Fig. MFN1 and MFN2 together with enhanced mitochondrial elongation have been identified in animals over-expressing mitochondrial OGG118. Figure XI, XII, XIII, and VII (4 h pre-PQ) also elevated MFN1 protein content after 48 h of PQ (0.6 mM) exposure. The highest levels of MFN1 protein were obtained with compound VII (25 μM), about 106% and 49% above basal and PQ levels, respectively. These data above are indicative of OGG1 mediated mitochondrial re-organization.Application of the OGG1 activators; 20. In the present study, the protein content of DNA ligase III (LIG3), which may be a rate-limiting component of the BER pathway36, was measured. Figure VII resulted in values that exceeded those values in its absence. In a separate experiment, 0.6 mM PQ increased OGG1 content by 88.3% above baseline . In the presence of 0.6 mM PQ, two different chemotypes, compounds VII (30 µM) and XI (50 µM) increased OGG1 content by 142.3% (p = 0.0056) and 227.4% (p = 0.002), respectively, above that achieved with PQ alone. The previous findings described above together with those found in the present investigation suggest that enhanced OGG1 activity by an as yet unknown mechanism enhances the protein content of members of the BER pathway for mtDNA repair.Excessive OGG1 activity could lead to the formation of apyrimic/apyrimidinic (AP) sites in the absence of sufficient participation by AP endonuclease 1 (APE1) and other downstream members of the BER pathway. Previous data using adenovirus to overexpress mitochondrial OGG1 in H9C2 cells exposed to oxidative stress showed that mRNA expression of BER members APE1 and DNA polymerase-γ was significantly upregulated by about twofold in the transduced cells compared to control onesDNA damage arising from ROS, which are a natural by-product of mitochondrial respiration, has long been considered to be a contributing factor to aging and its accompanying diseases. The magnitude of ROS exposure in human cells each day necessitates robust repair systems particularly within the mitochondria. As such, BER and specifically OGG1, have received considerable investigative attention. The present work expands on mechanisms of enhanced OGG1 catalysis for removal of 8-oxo-dG from DNA. Using small molecule OGG1 activators, we also demonstrate potential advantages for mitochondrial function during oxidative assault.24. The evidence for this unique mechanism  to free enzyme, further supports an allosteric mechanism of OGG1 action.Product-assisted catalysis was first proposed to describe enhancement of OGG1 catalysis37. Positive feedback regulation of OGG1 by 8-oxoGua could be beneficial during oxidative stress. It has been argued that OGG1, although bifunctional, is mainly responsible for catalyzing the glycosylase reaction in the intracellular context that produces an abasic DNA product under normal physiological conditions38. Allosteric activation of OGG1 AP-lyase activity mediated by the product of the OGG1 glycosylase reaction and its analogs, as shown herein, may provide a positive feedback regulation mechanism to enhance BER in cells undergoing repetitive oxidative stress. When 8-oxo-dG accumulates, it may allosterically bind to OGG1 following excision and enhance AP-lyase functionality to accelerate BER. In turn, cells can accommodate and, over time, adapt to a more hostile environment. The content of OGG1 in the nucleus, perhaps at the cost of mitochondrial dysfunction, increased in cultured cortical neurons following short periods (4 h) of oxidative stress39. Therefore, the notion of a product-mediated allosteric activation of OGG1 offers a mechanistic explanation of the OGG1 AP-lyase catalysis but also provides a plausible mechanism by which cells respond to oxidative stress.Feedback regulation, either positive or negative, is an efficient mechanism for cells to adapt to fluctuations in dynamic biochemical pathways in response to an ever-changing cellular environmentKm, an increase in Vmax, or a combination of the two as the result of activator binding to an allosteric binding site on the enzyme. The α value of ~ 1 determined from the initial rate analysis of OGG1 in the current study indicates that the allosteric activation of OGG1 by 8-oxoGua analogs , the chemical step itself and product dissociation40. The increase in Vmax of OGG1 by 8-bromoGua arises from acceleration of the chemical step40. An activation level of more than tenfold by 8-bromoGua has been reported previously24 which is consistent with the β-value of 36 , which is vitally important for energy production. The OGG1 activators attenuated this outcome and as shown here, preserved ATP levels. Moreover, the PQ-stimulated ROS occurred concomitantly with loss of mitochondrial membrane integrity and mtDNA damage in human A549 cells. Damage to mtDNA was diminished by two of the OGG1 activators. The direct application of H2O2 to a different cell type, i.e., MEF cells, decreased mtDNA copy number (CN) and its recovery was accelerated by the presence of the OGG1 activator, compound VII. DNA strand breaks, particularly mtDNA, lead to fragmentation and expulsion to elicit inflammation through several pathways, such as NF-κB translocation42. PQ also induced NF-κB translocation. The pretreatment of A549 cells with the OGG1 activators also offset cytochrome c translocation which precedes apoptosis. Although the TOM20 results did not indicate enhancement of mitochondrial density, the greater DRP1 and MFN1 protein levels in the presence of the OGG1 activators suggest heightened mitochondrial re-organization to preserve mtDNA integrity. The increased LIG3 and OGG1 content in response to PQ and the OGG1 compounds likely promotes improved BER activity to assure efficient repair of the damaged nucleobase. Lastly, cellular efficacy of these diverse “tool” compounds may be influenced by molecular weight, size, charge, lipophilicity and chemical stability in saline media.Some of the small molecule OGG1 activators described in the current work were previously shown to protect cells from accumulating PQ-induced 8-oxo-dG in mtDNAMth1 and Ogg1, 8-oxo-dG accumulated in mtDNA which was accompanied by a loss of MMP and mitochondrial function43. OGG1 siRNA rapidly decreased α-OGG1, which has a targeting sequence for mitochondrial import, coincident with progressive deterioration of MMP with menadione-induced mitochondrial superoxide production44. Respiratory capacity was, however, unaltered using OGG1 deficient (KO) mice45. Decreased mitochondrial function was unrelated to mass and due to fragmentation, suggesting that α-OGG1 is “essential to mitochondrial morphology and function after exogenous oxidative stress”44. Our previous investigation27 and the present one, are consistent with this perspective.The most prominent mitochondrial function is energy production, which is intimately dependent upon MMP. Recent studies suggest that OGG1 influences MMP. In cultured cortical neurons isolated from mice lacking the genes, 46. Single-stranded breaks in both genomes precedes cell death whereas breaks in mtDNA provoke its depletion. In turn, the loss of mtDNA CN lowers energy production to compromise cell viability34. By contrast, overexpression of mitochondrial OGG1 lowers mtDNA 8-oxo-dG levels with fewer mtDNA deletions and fragments with improved CN to support expression of mitochondrial encoded proteins, preservation of MMP and reduce apoptosis20. Sufficient CN is absolutely necessary for biogenesis of key oxidative phosphorylation proteins47. Depletion of CN impedes mtDNA replication47, while the persistence of abasic sites blocks replication48. Heteroplasmy and low levels of absolute CN may influence the onset of human age-related diseases49, whereas high levels of wild type mtDNA can offset deleterious pathogenic mtDNA mutations50 and support organ function51. Thus, the protection against PQ-induced mtDNA damage and the accelerated recovery of CN post-H2O2 exposure by our OGG1 activators may provide a therapeutic means to slow disease pathology.The accumulation of 8-oxo-dG lesions within DNA triggers serial events that are deleterious to mitochondrial function and cell viability. Menadione-induced oxidative stress in OGG1 KO cells provokes rapid cell death by two separate mechanisms, i.e., poly-ADP-ribose polymerase-dependent or calpain-mediated52, suffer greater levels of ischemic cortical infarct39 and with aging display a Parkinson’s disease phenotype53. Moreover, the KO mice have greater high fat diet (HFD)-induced adiposity with metabolic syndrome features54 and reduced gene expression of Krebs’ cycle components. By contrast, transgenic mice that overexpress mitochondrial OGG1 have lower levels of 8-oxo-dG within white adipose tissue, improved whole body energy expenditure and attenuated adiposity18. Mitochondria of OGG1 transgenic mice are elongated with increased expression of mitochondrial encoded genes with reduced levels of NF-κB. The latter phenotype indicates that enhanced OGG1 activity within mitochondria improves their function when stressed.The metabolic and functional ramifications of OGG1 deficiency (KO) or its overexpression, particularly within mitochondria, have been examined in several murine models of disease. As compared to wild type mice, OGG1 KO mice have 20-fold more 8-oxo-dG in their mtDNA27 and current findings demonstrate that OGG1 represents a promising drug target for discovering small molecule activators to treat oxidative stress-related diseases. Potentially contributing to these diseases are genetic and environmental factors that lower OGG1 efficacy. For example, activity by the loss of function OGG1 Ser326Cys variant, which is associated with Alzheimer’s Disease55 and all-cause mortality56, is improved by our small molecule OGG1 activators27. ROS-mediated oxidation of OGG1’s multiple cysteines or exposure to cadmium also lowers OGG1 catalysis58. Independent confirmation that small molecules can enhance OGG1 activity59, albeit by a separate mechanism than that described herein, i.e., via polynucleotide kinase phosphatase, strongly indicate that OGG1 is druggable. It is noteworthy, however, that some pro-inflammatory conditions may benefit from OGG1 inhibition61. The OGG1 inhibitor, TH5487, prevents TNF-α-induced OGG1 interactions with nuclear DNA to decrease NF-κB binding62. Oxidized mtDNA within mitochondria, if not readily repaired by OGG1, fragments and exits the organelle to initiate inflammation63. Nonetheless, allosteric activation by small molecules is favorable to drug discovery. The further optimization of these early, tool compounds toward selective, orally available, and potent small molecule OGG1 activators along with comparison to their inhibitory counterparts will promote greater understanding of their therapeutic potential.Collectively, our previousUnless otherwise stipulated, chemicals including 8-bromoGua, 8-aminoGua, and Benzonase® nuclease were purchased from Sigma-Aldrich , Roche  or for 9-deazaguanine (9-deazaGua), Santa Cruz Biotechnology . The purity of 9-deazaGua was > 99.5% as determined by LCMS  were lysed in buffer A  supplemented with Benzonase® nuclease and EDTA-free, complete, protease inhibitor cocktail tablets. The lysate was passed twice through an Avestin pressure drop homogenizer (12 kpsi). Cell debris was removed by centrifugation  and the supernatant mixed with Ni–NTA agarose resins equilibrated with buffer A. The mixture was rotated (2 h), centrifuged , resuspended in buffer A and packed into a XK26 column, which was then washed with buffer A until A280 reached baseline. The captured OGG1 protein was eluted with 0–50% buffer B (buffer A + 500 mM imidazole) gradient in buffer A. The eluted fractions were analyzed by SDS–PAGE. Fractions containing protein were pooled and concentrated using a 10-kDa molecular weight cut-off centrifugal concentrator. The tags of the OGG1 protein were removed by His6-Tev protease (enzyme: substrate ratio = 1:100) followed by passage through a Ni–NTA agarose column. Tag-free OGG1 was collected, concentrated, and injected into a Superdex 200 26/60 gel filtration column equilibrated with buffer A. The eluted protein was detected by a 4–12% gradient SDS–PAGE. Human OGG1 fractions were pooled, aliquoted and stored in buffer B plus 400 mM NaCl at − 80 °C. Both the full length (1–345 amino acids) and truncated (12–345) OGG1 proteins were prepared to 99% purity without DNA contamination measured by gel densiometry with chromatograms at an A280/A260 ratio. Both proteins were tested using a gel-based assay and the fluorescent kinetic assay described below, with comparable activity, however the truncated form was used in these studies. It has been reported that the OGG1 β-isoform in situ lacks glycosylase activity but protects mitochondrial function as an aconitase chaperone protein64.As described previously28 using an 8-oxodG-containing hairpin oligodeoxyribonucleotide  in water (100 μM) which was annealed  by heating (90 °C X 5 min) and cooling (4 °C at 0.05 °C/sec) the sample. All reactions  were run in Greiner 384 black low volume plates . The annealed substrate plus or minus activator was mixed with enzyme and the evolving fluorescence of each well was measured in kinetic mode . Initial rates were calculated from the linear phase and the molar concentration of the reaction product from a FAM standard curve.An assay was constructed based on published methodsν = the initial rate in the presence of an activator, Emax = maximum activity, EC50 = concentration of activator at which the enzyme displays half-maximal activity, B = background, and n = the Hill coefficient. Several Hits were assessed for binding to OGG1 using surface plasmon resonance . For representative values of compounds used in cell-based studies, see Supplementary table Compounds (> 2 million from the GSK collection) were dispensed (0.06 μl/well in DMSO) in 1536 Greiner black low volume plates. The 8-oxodG containing hairpin DNA substrate in 3 μl assay buffer was added to each plate well followed by addition of 3 μl enzyme. The concentration of the reactants in the compound screening protocol were: OGG1 = 10 nM, substrate A = 100 nM, and compounds were added at 20 µM. The level of DNA cleavage was: < 5% for OGG1 alone and about 20% for OGG1 + Hits or 8-bromoGua. Compounds with activation > 20% relative to that obtained with 500 μM 8-bromoGua, which was defined as 100%, were classified as potential Hits. After incubation (22 °C X 40 min), fluorescence intensity was determined . Concentration responsiveness was evaluated with these hits followed by a least squares analysis according to Eq. (6 (99.9 atom %D) and 20% D2O (99.9 atom %D). Line broadening of a single fluorine-containing OGG1 activator  upon binding to OGG1 (0–24 μM) was monitored at 564.17 MHz by 19F-NMR . Line narrowing resulting from displacement of the bound fluorine-containing compound was also measured in the presence of a potential competitive displacement compound e.g., compound VII (100 μM). 19F-NMR spectra were recorded using a 5 mm fluorine probe, regulated at 20 °C and the following parameters: 128 K complex data points, sweep width of 139,860 Hz, acquisition time of 0.469 s per scan, 384 scans and a relaxation delay of 1.85 s. Prior to Fourier transformation, sensitivity and resolution were enhanced by applying an exponential line broadening of 1 Hz over the first 128 K data points and zero filled to 512 K data points. Spectra were analyzed using in-house data analysis software.Samples were prepared in 20 mM Tris–HCl, pH 7.5, containing 150 mM NaCl, 0.1 mM TCEP, 2% DMSO-D65 using the published OGG1 crystal structure66. The simulation was carried out with a box of 80% water and 20% probes 24. An equilibration phase of 1.5 ns was followed by a 40 ns simulation. The threshold for a site to be a potential binding “hot-spot” was set to < -1 kcal/mol. Modeling small molecule interactions with OGG1 was accomplished using GlideXP67 as implemented in the 2017–4.11 Schrodinger Suite with OPLS200568 forcefield. OGG1 from the crystal structure24 was protonated (pH 7.4), whereas missing rotamers and contiguous gaps less than 5 amino acids were rebuilt using Prime69. Crystallographic waters greater than 5Å from the nearest sidechain were removed. For docking experiments, protein grids centered at each hot-spot were prepared  and a rigid receptor was used. Ligands were protonated and tautomerized to pH 7 (± 2) with those having a probability > 5% kept as distinct entities for docking with up to 500 conformations/ligand. Each minimized docked pose was visually inspected. Those with nonsensical interactions or obviously strained geometries were removed. Docking-induced strain was calculated by assessing the distance between global and local minima (OPLS2005).To identify potential small molecule binding sites, mixed probe molecular dynamics were carried out as previously described27 using complete Ham's F12 medium with 10% heat-inactivated fetal bovine serum (FBS), and GlutaMAX™ supplement  in O2:CO2; 95:5% at 37 °C. For sub-culture, the cells were treated with Trypsin/EDTA , counted , and seeded . Oxidative stress was invoked by paraquat  at varying concentrations (0.01–3 mM) and incubation periods (refer to figure legends). PQ was dissolved in Dulbecco’s phosphate buffered saline and stored in aliquots  until use when they were thawed and diluted in cell culture medium. Unless stated otherwise, OGG1 activators were incubated with cells for 4 h prior to treatment with PQ. All measures were performed in triplicate on three independent occasions unless stated otherwise. For specific details describing the immunofluorescence staining and quantification please refer to the Supplemental materials.Type II alveolar epithelial cells, A549 , were grown to sub-confluence as described previously2 at 37 °C. For sub-culture, cells were trypsinized , counted using a cytosmart cell counter , and 100,000 cells were plated per well in 12-well plates. Cells were cultured for 24 h prior to a 4 h treatment with 30 µM compound (in DMSO) or vehicle. For hydrogen peroxide (H202) treatments, the stimulus concentration was determined by the absorbance at 240 nm (1.31 = 30 mM H202) and adjusted in calculations. Cells were washed with PBS and exposed to 400 µM H202 in DMEM without FBS for 15 min. At that time, H202 was neutralized by the addition of an equal volume of DMEM with 2X FBS, all media removed, cells washed with PBS, and warm DMEM with FBS added for the indicated recovery time. To collect cells, plates were washed with PBS, trypsinized and collected by centrifugation prior to flash freezing in liquid nitrogen. All samples were collected and kept at -80˚C until analysis for mtDNA determination. MEFs were kindly provided by Dr. Mark Gladwin, University of Pittsburgh Medical Center. All work regarding the collection of the MEFs were conducted in accordance with Institutional Policy on the Care, Welfare and Treatment of Laboratory Animals and were reviewed by the Institutional Animal Care and Use Committee using an ethical review process at the institution where the work was performed.Mouse embryonic fibroblasts (MEFs) were grown in Dulbecco’s Modified Eagle Media  with 10% FBS at 5% CO70. After freeze–thaw of resuspended DNA, total DNA was quantitated by AccuBlue®  and assayed for relative nuclear DNA (nDNA) levels by quantitative PCR as described71. Samples were adjusted to be within + /- 1 Cq value and assayed in duplex for both mtDNA  and nDNA (HDAC1) against a pooled sample standard curve. Both nuclear and mitochondrial targets were plotted and validated to be in the linear range. Furthermore, ∆Cq values were plotted to assess the effect of dilution on relative mtDNA levels. With these validations in place, we determined the relative mtDNA levels .MEFs pellets were rapidly thawed and digested with SDS/proteinase K as previously described72. Long-chain PCR was conducted using the 8.9 kilobase mtDNA fragment and primers  with accession numbers 14841 and 5999; and for short-chain PCR using the small mt fragment and accession numbers 14620 and 1484134. The PCR product was quantified using PicoGreen™ . Fluorescence intensity of the long-chain was subtracted from that of the short-chain and expressed as a percent of the untreated control.For mtDNA damage, A549 cells were lysed using the Cells-to-Ct kit . DNA was isolated, quantified and followed by long and short chain PCR reactions according to published methodsAdenosine triphosphate (ATP) and adenosine diphosphate (ADP) levels in A549 cells cultured in 6-well plates were measured following treatment with or without PQ  using commercially available kits . Samples were collected from separate plates for each metabolite, lysed, stored at − 80 °C for analysis. Perchloric acid (4 M) extracts were prepared, neutralized and fluorescence intensity  was monitored using an Envision Multi-label plate reader . ATP and ADP levels were normalized to protein concentration .p < 0.05.Data represents mean ± SEM unless stated otherwise. Initial rates were analyzed using a least-squares analysis. Comparisons were made by ANOVA with Dunnett’s post-hoc analysis or one-or two-way students’ t-test using GraphPad Prism. Minimal statistical significance was set at Supplementary Information.


Cervical 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.The 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.In 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.Various 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. Cervical cancer is a common female malignant tumor. According to the statistic results in 2020 all over the world , cervicaHuman papillomavirus (HPV) is a spherical and non-enveloped virus with a double-stranded circular DNA genome, showing strong squamous epithelium-like characteristics to infect damaged epithelial cells . More thIt has been proved that long-term and persistent infection of HR-HPV is closely related to the occurrence and development of cervical cancer . HoweverVirus variations may cause differences in immune response and oncogenic potential . There aIn this study, samples of cervical exfoliated cells were collected from the female patients with possible HR-HPV infection who underwent routine cervical screenings at the Department of Gynecology of Jingzhou Hospital from Spe. 2019 to Oct. 2021. All the samples were stored at 4 °C, and DNA was extracted within 1 day. Informed consent of patients had been obtained, and the study subjects’ privacy was protected before sample collection.4copies/mL; coincidence rate of positive reference: 100%; coincidence rate of negative reference: 100%; no cross-reaction with HPV-11, -6, -42, -43, -44, -73, -26, -53 types, Chlamydia trachomatis, Ureaplasma urealyticum, Neisseria gonorrhoeae, Mycoplasma hominis, Mycoplasma genitalium, Candida albicans, HIV-1, group B streptococcus). Finally, DNA extraction products of single positive samples of HPV-58 were selected and stored at − 80 °C for the follow-up experiments.Based on the method of magnetic beads, DNA was extracted according to the instruction of the nucleic acid extracting kit . 4 μL extraction products were used for HPV typing as a PCR template. According to the instruction of the HR-HPV typing test kit , DNA extraction products were detected through the method of real-time quantitative PCR, including 15 types of HR-HPV , with advantages of excellent sensitivity, specificity and typing accurately  (Takara), 2 μL dNTPs (Takara), 0.125 μL Taq polymerase (Takara), and 1.25 μL forward and reverse primers (20 μM) (Sangon Biotech). The amplification conditions of E6 were initial denaturation at 95 °C for 3 min, followed by 35 cycles of 94 °C for 45 s, 58.2 °C for 45 s, 72 °C for 1 min, and the 72 °C for 10 min. The E7 amplification conditions were initial denaturation at 95 °C for 3 min, followed by 35 cycles of 94 °C for 45 s, 62.1 °C for 45 s, 72 °C for 1 min, and the 72 °C for 10 min. The PCR products were analyzed by electrophoresis in 2.5% agarose gel stained with GelRed nucleic acid dye (Biotium), and then sent to Sangon Biotech for sequencing.The E6 and E7 genes were amplified from HPV-58 single positive samples. According to the reference sequence provided by GenBank (D90400), software Primer Primer 6.0 was used to design sequence-specific primers of HPV-58 E6 and E7 genes following primer design principles. All primers were synthesized by Sangon Biotech (Sangon Biotech (Shanghai) Co., Ltd.). As followed, E6 F:5′- TTGGGTCACATTGTTCATG-3′, E6 R:5′-TCATAGCAGAATAGGTCAGT-3′; E7 F:5′- ATTTCGGGTCGTTGGACAGG-3′, E7 R:5′- TCTGTACCACTATCGTCTGCTGTT-3′. 25 μL reaction volume included 2 μL HPV DNA extraction, 15.875 μL ddHThe nucleotide variants and amino acid substitutions were analyzed by comparing all sequencing results with HPV-58 reference sequence (D90400) via the DNAstar and NCBI blast.The Neighbor-Joining tree of HPV-58 E6 and E7 sequences was constructed by MEGA 11 software, Kimura 2-parameter model was used, and the Bootstrap replications was set at 1000. The following reference sequences collected from GeneBank sequences database were used to construct the phylogenetic branches: D90400 (A1), HQ537752 (A2), HQ537758 (A3), HQ537762 (B1), HQ537764 (B2), HQ537774 (C), HQ537768 (D1), HQ537770 (D2) .http://bioinf.cs.ucl.ac.uk/psipred/) was used to predict the secondary structure of E6 and E7 amino acid sequences. The selective pressure was analyzed by PAML X software. The non-synonymous and synonymous nucleotide divergence (dN/dS) was calculated by Codeml program. Using the Bayes Empirical Bayes (BEB) analysis, the sites with the posterior probability ≥ 0.95 were identified as the positively selected sites.The online server PsiPred (We predicted linear B cell epitopes of E6 and E7 proteins by online software ABCpred (ABCpred submission page (iiitd.edu.in)) according to the default parameters. The higher the predicted score, the better the affinity of the epitope.A total of 18,266 samples were collected in Jingzhou Hospital, Yangtze University from Sep. 2019 to Oct. 2021. The overall prevalence of HR-HPV infection was 18.58% (3394/18266), with 14.62% (2671/18266) of participants being positive for a single HPV type, and the age group with highest infection rate was 58 to 67 years (32.43%). The detailed results are shown in Table There were 319 single positive samples of HPV-58, and some samples were failed amplified due to the low viral load or degradation of viral nucleic acid. Finally, 172 entire E6-E7 genes were successfully amplified and further analyzed. Compared with the reference sequence D90400, the prototype accounted for 18.02% (31/172) of the total sequencing samples, and the remaining 141 sequences were divided into 21 different variant groups named 58HB01-58HB21, which were submitted to the GenBank database and got the accession numbers were OL989036 to OL989056.A total of 22 single nucleotide polymorphisms (SNPs) were observed in E6-E7 sequences. In E6 sequences, 10 SNPs were found, including 7 synonymous variants and 3 non-synonymous variants. The prevalent variants were A388C (K93N) (81/172) and C307T (57/172). In E7 sequences, 12 SNPs were observed, including 3 synonymous variants and 9 non-synonymous variants. T744G (104/172), G761A (G63D) (51/172), G694A (G41R) (47/172) and T803C (V77A) (46/172) were common variants. G261A (R51K), A310G, A340G and T407G in E6 sequence and A597G in E7 sequence were novel variants. Two non-synonymous variants (R51K and L100V) occurred in E6 sequences encoding the alpha helix, two non-synonymous variants (R9K and V77A) were found in E7 sequences encoding the strand. No insertion or deletion was detected in any sequence. The detailed results are shown in Table The phylogenetic tree was constructed with 21 unique HPV-58 E6-E7 sequences and 8 sub-lineage reference sequences by MEGA 11 software. The phylogenetic tree is shown in Fig. The variable dN/dS rate ratios were tested among the various lineages using the PAML X. In HPV-58 E6 sequence, there was no positively selected site. In E7, codon site G63 was identified as the positively selected site. All the results are shown in Table To evaluate the influence of HPV-58 E6 and E7 sequence polymorphisms on the host immune response and recognition, online server ABCpred was used to predict the potential B cell epitopes of E6 and E7 proteins. The epitope prediction scores should be greater than 0.85. The results are shown in Table Persistent infection of HR-HPV is the main factor for the occurrence and development of cervical cancer. The E6 and E7 proteins encode by E6 and E7 genes are the main carcinogenic proteins and play an important role in the cancer progression. HPV variants may be linked to the evolution of human groups to get an intrinsic geographical difference in prevalence and infections. The variants analysis of HPV E6 and E7 gene from clinical samples is of great significant for further study on vaccine and cervical cancer therapy in local area.In this present study, we found that HPV-52, 58, and 16 types were prevalent types in Jingzhou area. The infection rate of HPV-58 was 17.00%, ranked second. This unusual high prevalence has also been reported in Chinese populations in Xi’an, Guangxi, and Chongqing –22.The pThe results of E6-E7 sequencing showed that the sequence variability of E7 sequence (4.0%) was higher than that of E6 sequence (2.2%), suggesting that E6 may be a more suitable target for HPV therapeutic vaccine than E7. In E6 gene, C307T and A388C (K93N) were common variants. In E7 gene, T744G was most prevalent synonymous variant, G761A (G63D), G694A (G41R), and T803C (V77A) were common non-synonymous variants. There were four non-synonymous variants located in the sequences encoding the secondary structure of E6 and E7 proteins, may cause changes in oncoprotein folding and function, led to the differences in their ability to interact with tumor suppressor proteins, thus affecting the pathogenicity of HPV-58 . Among tHPV-58 type has four lineages of A, B, C and D. Lineage A is significantly more frequent in Asia than in America, lineage B is mainly distributed in America, and lineage C and D seem to be more frequent in Africa . In thisAt present, all HPV vaccines on the market  are prophylactic vaccines based on L1 protein immunogenicity, which play an effective role in preventing HPV infection by producing specific neutralizing antibodies. However, the vaccines do not prevent cancer development in individuals who are already infected . TherefoThis study analyzed the positive selection of HPV-58 E6 and E7 sequences in Jingzhou area. The main feature of positive selection is that it causes an unusual repaid increase in the frequency of alleles, which allows a species to adapt quickly to environmental changes . The posIn this study, we analyzed the gene polymorphism of HPV-58 E6-E7 sequence and amino acid substitution of E6-E7 proteins in Jingzhou area, constructed a phylogenetic tree to analyze the lineage characteristics, and predicted the B cell epitope to clarify the regional specificity of HPV-58 gene variation. Some new variations and B cell epitopes were observed in this study, and we also got a general knowledge of the gene polymorphism of HPV-58 in Jingzhou area. This study can supplement the knowledge of gene diversity of HPV-58 in central China, and provide an experimental basis for subsequent studies on epidemiology, evolution, pathogenicity and therapeutic targets of HPV-58.