Methods. In this cross-sectional study, data were collected by the purposely constructed questionnaire. A questionnaire composed of the demographic items and items related to the awareness and knowledge about Human papillomavirus. The questionnaire was constructed after a series of discussions between the panel of experts. This panel was composed of a subject specialist, researcher, and language expert. The Cronbach alpha of the questionnaire was calculated. The study will be conducted in the Al Qunfudhah. Results. The Cronbach alpha of the questionnaire was 0.72. Out of a total of 550 respondents, with a mean (SD) age of 47.5 (11.5), the female respondents were 167 (30.4%) while male were 383 (69.6%). 20.5% of the respondents (out of 550) were having awareness of HPV. Implications. Knowledge of HPV-related oral cancer is critical, and it is advised to be taught as part of dental students' basic curriculum and clinical training. This problem can be solved by better educational training programs. Knowledge of HPV-related oral cancer is critical, and it is advised to be taught as part of dental students' basic curriculum and clinical training.With 14 million new infections each year, the human papillomavirus (HPV) is the most common sexually transmitted infection (STI) among both men and women in the United States (US). Infections with the human papillomavirus (HPV) are responsible for a considerable portion of the global cancer burden. HPV-related oral malignancies are on the rise around the world, according to epidemiological studies. To provide accurate advice to their patients, dental practitioners require thorough, up-to-date HPV-related knowledge.  With 14 million new infections each year, the human papillomavirus (HPV) is the most common sexually transmitted infection (STI) among both men and women in the United States (US). Infections with the human papillomavirus (HPV) are responsible for a considerable portion of the global cancer burden. HPV-related oral malignancies are on the rise around the world, according to epidemiological studies. To provide accurate advice to their patients, dental practitioners require thorough, up-to-date HPV-related knowledge.TSeropositive men are more likely to acquire penile and anal cancer after contracting HPV, and other studies found that HIV–HPV seropositive people are 60 percent more likely to get cancer than healthy people. 10 heterosexual HPV-positive persons, on the other hand, can transfer the virus to their female partners, which is linked to cervical cancer in these women. Skin-to-skin or skin-to-mucosa contact is the most common way for HPV infections to spread. More than 200 HPV kinds have been identified thus far [The epidemiology of HPV in Saudi Arabian women is poorly understood, and there are few publications in this community about HPV prevalence, detection, and genotyping, as well as attitudes about screening.In 2018, a study in Saudi Arabia discovered that oropharyngeal cancer is more common in men than in women. According to the findings, there were 12 new cancer cases each year in men, with a crude incidence rate of 0.1, an age-standardized incidence rate of 0.1, and cumulative risk (%) of 0 at 75 years old. The annual number of new cancer cases in females was 6, with a crude incidence rate of 6 and an age-standardized incidence rate of 0 percent [Based on their carcinogenic potential, HPVs are categorized into two categories: high risk and low risk. High-risk strains include HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82 [Oncogenic strains of the HPV are associated with at least six different cancers including oropharyngeal, anal, cervical, vaginal, vulvar, and penile cancers –9. The lHPV has been discovered as an oncogenic driver in HNC, according to a study conducted in Canada in 2016, and the prevalence of HPV + HNC, particularly HPV + OPC, is growing . In 2005Fortunately, three HPV vaccinations have been developed to protect against up to nine different strains of HPV . The quaBecause of a lack of proper awareness and understanding regarding HPV-related infections and malignancies, we need to enhance the knowledge According to recent studies conducted in Florida and Italy. Very few respondents were aware of the presence and availability of an HPV vaccine , 16. OnlIn a similar study conducted on physicians in Saudi Arabia in 2017–2018, it was discovered that physicians are only partially aware of the evident dangers of HPV infection to the Saudi population. Physicians, on the other hand, frequently reported modest levels of concern about HPV infection. Lack of information about the vaccine and concerns about vaccine safety and cost were cited as reasons for rejecting the vaccine, demonstrating a widespread unfavorable attitude about the HPV vaccine that may underplay clinicians' role in prescribing immunization for their patients [Due to the paucity of information available on the extent of people's awareness and knowledge of HPV disease and vaccines in Al Qunfudhah and the lack of published research on this point, in this study, we aim to assess the extent of people's awareness and knowledge of the HPV diseases, methods of transmission, vaccines, and related cancers.To assess the awareness of HPV infection and oropharyngeal cancer in people in the Al Qunfudhah region, Kingdom of Saudi Arabia.To estimate the knowledge and attitude of HPV infections, related cancer, and vaccination in people in the Al Qunfudhah region, Kingdom of Saudi Arabia.To calculate the prevalence of HPV infection in people in the Al Qunfudhah region, Kingdom of Saudi Arabia.To assess the effect of education on the participant's knowledge and awareness about HPV infections.In this cross-sectional study, data were collected by the purposely constructed questionnaire, a questionnaire composed of the demographic items and items related to the awareness and knowledge about the Human papillomavirus. A questionnaire was constructed after a series of discussions between the panel of experts. This panel was composed of a subject specialist, researcher, and language expert. Cronbach alpha of the questionnaire was calculated. The study was conducted in the Al Qunfudhah region of Saudi Arabia. The questionnaire included items related to the awareness of HPV, demographic variables, the vaccine, and other items.t-test and chi-square test was used at 5% level of significance. Data were collected from the patients who visit the Primary Health Care Center at Al Qunfidah city of Saudi Arabia and from the general public of A Qunfidah city of Saudi Arabia through an electronic version of the questionnaire. Ethical approval was obtained from King Khalid University, Saudi Arabia. The study duration was from January 2021 to April 2021.After the collection of data, data were coded and entered in the SPSS ver. 20 software for descriptive statistics , to measure the significance differences The Cronbach alpha of the questionnaire was 0.72. Out of a total of 550 respondents, with mean (SD) age of 47.5 (11.5), the female respondents were 167 (30.4%) while males were 383 (69.6%). We observed that 20.5% (out of 550) respondents were having awareness of HPV. We observed a significant difference while comparing gender with awareness questions. Further, 29% (out of 113 respondents) agreed that HPV vaccination can prevent oropharyngeal cancer.In Out of 113 respondents (those who were aware of HPV) opted that HPV can cause HIV aids (24% agreed), 49% had no idea, and 27% disagree .In this study, our major aim was to assess the awareness and attitude towards HPV infections, diseases, and vaccination associated with HPV. In the United States, roughly, 42% of persons aged 18 to 59 have genital human papillomavirus (HPV) infections, while about 7% have oral HPV infections , 22. AccHPV is a group of DNA viruses that infect basal epithelial cells and cause benign and malignant diseases of the skin, anogenital mucosae, and upper aerodigestive tract mucosae. 4–6 HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 59 have been classified as carcinogenic, while HPV68 has been classified as possibly carcinogenic, based on epidemiological research and mechanistic evidence. HPV is the cause of nearly all cervical cancers, as well as a significant number of other anogenital and oropharyngeal malignancies. Understanding the cancer burden caused by HPV can help to improve HPV vaccine and HPV-based cervical screening programs. In our study, we found that a reasonable quantity of the respondents agreed that HPV can cause different types of cancers –28.On the role of HPV in cervical cancer, there is significant epidemiological evidence . In our study, a significant number of the respondents agreed to have a vaccination for HPV as the study stated that. Vaccine efficacy against high-risk HPV types (HPV) has been found to range from 90 to 99 percent in preventing cervical cancer in numerous clinical trials and systematic reviews around the world (CIN). These assessments also provide ample evidence of the vaccinations' safety .Many underdeveloped nations have a low uptake of HPV vaccination due to factors such as a decreased perceived risk of cervical cancer, a lack of vaccine availability, and exorbitant costs. However, one of the most critical deciding reasons for low vaccination uptake is a lack of proper knowledge regarding the role of HPV in the development of cervical carcinoma and the HPV vaccine , 31.According to numerous studies conducted among physicians and nurses, the most crucial aspects evaluated by clinicians when considering the HPV vaccine were effectiveness and side effects/safety. Government recommendations and expenses were also taken into account. The importance of clear government advice and cost were also examined. Many of these studies underlined the need for additional information regarding the virus, cervical cancer, and vaccination to boost people's readiness to suggest it –35.The signs and symptoms of HPV are usually not visible, which is also stated in our study that the majority of the respondents were unfamiliar regarding the signs and symptoms of HPV .In our study, a reasonable volume of the respondents agreed that the HPV vaccination is a method for preventing oropharyngeal cancer which is also proof by the study stating that HPV VLP vaccinations of the current generation (HPV 16/18) have the potential to prevent >90% of HPV-positive oropharyngeal malignancies. Men and possibly women should be included in clinical trials evaluating the efficacy of HPV VLP vaccinations for the prevention of incident and chronic oral HPV 16 infection .In women who live in a place where both illnesses are common, cervical HPV infection is linked to HIV infection. Much research is needed to determine whether the reported link is causative. In our study, 24% of the population agreed that cervical HPV infection is linked to HIV infection.Education might play an important role in having HPV vaccination and avoiding cancers; in our study, university graduates having more desire to get vaccinations of HPV which is inconsistent with the findings of the study stating that to achieve immunization, educational programs aimed at health care practitioners, as well as cost-cutting initiatives and clear government rules, are urgently required .These findings imply that respondents have significant HPV-related knowledge gaps, which limit their ability to intervene in oropharyngeal primary prevention initiatives. The findings of this study recommend that HPV-related instructional content be included and standardized in school, college, and university curricula. This problem can be solved by better educational training programs. Knowledge of HPV-related oral cancer is critical, and it is advised that it should be taught as part of dental students' basic curriculum and clinical training. This research will serve as a foundation for future research and therapies. Medical students should do a follow-up comparative study.


Background and Objectives: To investigate the effect of infection with human papillomavirus (HPV) or Chlamydia trachomatis (CT) and HPV + CT coinfection on sperm quality, inflammation, and the state of oxidative stress (OS) in asymptomatic infertile men. Materials and Methods: Semen samples from 84 asymptomatic military infertile men were studied. The polymerase chain reaction (PCR) was used for the molecular detection of HPV and CT. Semen parameters were analyzed according to the World Health Organization guidelines. Inflammation was evaluated by an IL-1β, IL-6, and IFN-γ enzyme-linked immunosorbent assay (ELISA) and OS by the quantification of lipid peroxidation (LPO), 8-hydroxydeoxyguanosine (8-OHdG), and total antioxidant capacity (TAC). Results: A total of 81 of the 84 (96.4%) samples were positives for the pathogens, with 55/81 (68%) being positive for HPV, 11/81 (13.5%) for CT, and 15/81 (18.5%) for HPV + CT coinfection. Seminal parameters were affected in the infected groups, including pH increases above the normal range in all groups. An abnormal sperm morphology was observed in the HPV and HPV + CT groups. Higher cytokine levels were detected in the HPV group and the highest IL-1β level was found in the HPV + CT group. No cytokines were detected in the CT group. High LPO and 8-OHdG levels were found in all groups with a lower TAC. Comparisons between groups showed the highest OS state was observed in the HPV group. Conclusions: High HPV infection or coinfection (HVP + CT) in these infertile men suggest compromising male fertility by inducing a proinflammatory state and OS. Infection with CT suggests an alteration of the state of OS by promoting an alkaline pH. Chlamydia trachomatis (CT) are the most common sexually transmitted infections (STIs) worldwide [Infections with human papillomavirus (HPV) and orldwide ,2,3, whiorldwide  and may orldwide . Both paorldwide ,3. Notaborldwide ,5 and coorldwide ,6; howevorldwide , and canorldwide , lipid porldwide , and eveorldwide .Human semen contains cytokines that exert pleiotropic and redundant effects during the activation of innate, cellular, and inflammatory responses; in addition, the growth and differentiation of germ cells and functions of genital organs are regulated by proinflammatory cytokines such as IL-1β, IL-6, IL-8, TNF-α, and IFN-γ. Cytokines induced by infections, inflammation, and oxidative stress (OS) disrupt the male accessory gland function, ejaculate parameters, and sperm performance . MoreoveThis cross-sectional study included 84 male military service members with a mean age of 31.81 ± 5.71 years; this population was selected during infertility consultation between January and November 2019 in the “Hospital Militar de Especialidades de la Mujer y Neonatología” of the National Defense Ministry, México City. Inclusion criteria were men attending the hospital for conjugal infertility investigation who failed to conceive after one year of unprotected intercourse and who agreed to be subjected to sperm parameter analysis. Men under drug therapy and those with undescended testes, varicocele, and other structural abnormalities were excluded.g for 10 min to isolate seminal plasma and cell pellets were separated and stored at −20 °C until the time of analysis.Before semen collection and analysis, the men were asked to refrain from sexual intercourse for 3–5 days. Immediately after collection, the samples were liquefied at room temperature for up to 30 min; then, semen analysis was performed according to World Health Organization guidelines . The semChlamydia trachomatis. DNA was extracted from the semen samples using the DNeasy Blood and Tissue Kit . The HPV genome was detected by PCR using universal primers  [Detection of HPV and L1C1/C2) . AmplifiL1C1/C2) . The betThe seminal content of proinflammatory cytokines such as IFN-γ (range: 8–3000 pg/mL), IL-1β (range: 8–1000 pg/mL), and IL-6 (range:24–1500 pg/mL) was determined using a quantitative sandwich ELISA method. Standard curves were constructed for each cytokine according to the manufacturer’s instructions . Seminal cytokine concentrations are expressed as pg/mL of seminal plasma.v/v) was added. The final mixture was then heated for 30 min at 95 °C. After cooling to room temperature, the mixture was centrifuged at 3000× g for 10 min and absorbance of the supernatant was read in a spectrophotometer  at 532 nm. The results were expressed as nanomoles of MDA per milligram of protein. The protein levels were quantified by the Bradford protein assay using bovine serum albumin as standard.LPO was estimated in the semen samples by the measurement of malondialdehyde (MDA) using the thiobarbituric acid (TBA) method, with minor modifications . BrieflyDNA oxidation was evaluated by the formation of 8-hydroxydeoxyguanosine (8-OHdG), a ubiquitous marker of OS. The levels of 8-OHdG in 100 µL of seminal plasma were measured using the OxiSelect Oxidative DNA Damage ELISA Kit . The absorbance of each sample was determined in a microplate reader  at a wavelength of 450 nm and the levels of 8-OHdG were calculated from a standard curve. The minimum detectable concentration of 8-OHdG was 20 ng/mL and the maximum detectable concentration was 100 ng/mL.S-triazine (TPTZ) in 40 mmol/L HCl and with 1 volume of 20 mmol/L FeCl3.6H2O, and was warmed to 37 °C. Next, 30 µL of seminal plasma, 90 µL of distilled water, and 30 µL of each standard solution  were transferred to an Eppendorf tube (2 mL) and 900 µL of the FRAP working solution was added. The mixture was heated to 37 °C for 10 min. Finally, the absorbance values of blank, standard solutions, and samples were read in a spectrophotometer  at 593 nm. The results were corrected for dilution and are expressed as µmol FeSO4/L. All solutions were freshly prepared and immediately used. Measurements were performed in triplicate.The non-enzymatic TAC of seminal plasma was measured by the ferric reducing antioxidant power (FRAP) method ,17, withp value < 0.05 was considered statistically significant. All analyses were performed using the Statistical Package for the Social Sciences (SPSS)  and Microsoft Excel (Windows 10).Seminal HPV, CT, and HPV + CT coinfection are described as frequency (%). Statistical methods included mean and standard deviation (SD) for normal distribution and median (25th–75th percentile) for abnormal distribution. Specifically, the Mann–Whitney U test was used for comparison of two groups and one-factor analysis of variance (ANOVA) with post hoc Tukey or the Kruskal–Wallis one-way ANOVA was used for comparisons of more than two groups. A The study protocol was approved by the institutional review board of The “Hospital Militar de Especialidades de la Mujer y Neonatología” of the National Defense Ministry, México City (CONBIOÉTICA-09-CEI-015-20180924) and informed consent was obtained from all participants. All procedures were conducted in accordance with the Declaration of Helsinki and with the Official Mexican Standard (NOM-012-SSA3-2012).The results of the molecular assay demonstrated that 81 of the 84 samples (96.4%) were positives for the pathogens, with 55/81 (68%) being positive for HPV and 11/81 (13.5%) for CT and coinfection being detected in 15/81 (18.5%) as Seminal parameters, cytokines, and OS biomarkers were analyzed and compared between the infected groups: HPV, CT, and HPV + CT . An analThe highest cytokine levels were observed in the HPV group; however, IL-1β levels were higher in the coinfected group than in the HPV group and no cytokines were found in the CT-infected group. LPO and 8-OHdG levels were detected in all groups, in accordance with the lower TAC levels; nevertheless, the highest state of OS was found in the HPV group.The relevance of STIs such as those caused by HPV and CT as an etiological factor of male infertility remains controversial. Most studies report that these pathogens affect semen quality in asymptomatic infertile men ,2,18. SoA high prevalence of seminal HPV infection has been demonstrated by nested PCR in oligospermic and azoospermic men  .Trichomonas vaginalis, CT and HSV-1/-2 (9.6% each), highlighting that HPV + CT coinfection in the semen samples (7.1%) of asymptomatic sexually active young men (31.81 ± 5.71 years) can influence their reproductive health [This study highlighted the high prevalence of HPV infection (68%) and HPV + CT coinfection (18.5%), which is probably attributable to the specific population of military asymptomatic infertile patients analyzed who may be affected by other STIs. Other authors studying the Mexican army population found a high prevalence of seminal HPV in healthy soldiers (49%) and concluded that a risky sexual behavior, a large number of sexual partners, and anal intercourse increase the risk of acquiring and transmitting an STI . Additioe health .In this work, CT infection and CT + HPV coinfection were found in 13.5% and 18.5% of men, respectively, and the data were similar to other reports of men studied for infertility . HoweverLactobacillus in healthy men [The results suggest that semen quality was altered by the pathogens, as demonstrated by a pH increase. Recent studies suggest that infection with CT  and HPV lthy men . Lactobalthy men . The imbAccording to Damke et al., HPV semen infection may modify the semen volume decrease or increase) and augment pH (≥7.8), indicating altered proportions of the fluid secreted by the major sexual accessory glands  . ChangesThe negative impact of HPV or CT infection on sperm morphology is not well understood. It is hypothesized that morphological disturbances may result from the ability of the virus to bind to the spermatozoa head (through interactions between HPV capsid protein L1 and syndecan-1), as reported for HPV infection . AccordiPathogens can induce the secretion of proinflammatory cytokines and OS in the semen of asymptomatic infertile men. Cytokines play a central role in physiological and pathological processes in the male reproductive tract, innate and cellular immune responses, inflammation, growth regulation, germ cell differentiation, and intracellular signal transduction . FraczekThe overproduction of RONS or reactive oxygen species (ROS) impairs sperm function ,25. PossThe results suggest that infection with CT contributed to OS characterized by augmented LPO and diminished TAC levels, as reported previously ,28. HPV The high prevalence of HPV infection and HPV + CT coinfection detected in this study of infertile men, suggests a significant negative impact on male fertility, which could contribute to induce a proinflammatory state , particularly an increase in IL-1β in HPV + CT coinfection. HPV or HPV + CT could induce OS by an increase in LPO and 8-OHdG and a decrease in non-enzymatic antioxidant systems, promoting an alkaline pH, alterations in sperm morphology, and decreased sperm motility A of military asymptomatic infertile men.The results suggest that CT infection contributed to OS characterized by an increased LPO, decreased TAC levels, and increased seminal pH as seen in The main limitation of this study included its cross-sectional design, due to it being a study of men attending the hospital for conjugal infertility investigation, and the small sample size.Thus, a case–control study involving a more significant number of participants is required (fertile and infertile men). Studies in progress that involve healthy fertile controls of general populations and military communities could confirm the evidence obtained here. The repetition of HPV screening in semen after 12 months would be necessary to determine the clearance or persistence of seminal HPV infection. The results highlight IFN-γ, IL-1β, IL-6, LPO, 8-OHdG, and TAC evaluation in seminal plasma for a comprehensive state of male infertility to improve therapeutic intervention in male partners of infertile couples and to prevent infections or diseases.


Based on the etiology, HNSCC has two main subtypes: human papillomavirus (HPV) -related and HPV-unrelated. HPV-positive HNSCC is more sensitive to treatment with favorable survival. Due to the different biological behaviors, individual therapy is necessary and urgently required to deduce the therapeutic intensity of HPV-positive disease and look for a more effective and toxicity-acceptable regimen for HPV-negative disease. EGFR amplification and PI3K/AKT/mTOR pathway aberrant activation are quite common in HPV-positive HNSCC. Besides, HPV infection alters immune cell infiltrating in HNSCC and encompasses a diverse and heterogeneous landscape with more immune infiltration. On the other hand, the chance of HPV-negative cancers harboring mutation on the P53 gene is significantly higher than that of HPV-positive disease. This review focuses on the updated preclinical and clinical data of HPV-positive and HPV-negative HNSCC and discusses the therapeutic strategies of different HPV status in HNSCC.Head and neck squamous cell carcinoma (HNSCC) is the 9 The HPV-positive disease occurs predominantly in the oropharynx p<0.001) Radiotherapy and chemotherapy are critical therapeutic ways to treat HNSCC, and they are even the gold standard for patients with local advanced stage or recurrence and metastasis disease. RTOG 0129, the most cited trial, reported that HPV-positive patients had better 3-year rates of overall survival .The clinical significance of HPV status indicates the prognosis but cannot change the clinical decision-making. According to the guidelines of NCCN 2020, surgery or radiotherapy alone can be considered for HPV-positive oropharyngeal carcinoma in the T1-2 N0 stage, surgery, radiotherapy, or concurrent radiotherapy and chemotherapy can be regarded as in T1-2N1 , while tumors in a later stage can consider surgery, concurrent radiotherapy, and chemotherapy or sequential radiotherapy and chemotherapy after induced chemotherapy. For radical radiotherapy, the equivalent biological dose (EQD2) is still required to 70Gy or above . However, because of the treatment sensitivity of HPV-positive oropharyngeal carcinoma, appropriately reducing the treatment intensity would be an ideal strategy Reducing the intensity of chemotherapy is another strategy for HPV-positive disease. RTOG 1016 indicates that cetuximab has no merit to improve prognosis As described above, HPV status contributes less to conventional treatment. As we know, cancer derives through the accumulation of genetic and epigenetic alteration in genes involved in a variety of signaling pathways and precipitates the cancer-associated phenotypes About 70% of HNSCC has an epidermal growth factor receptor (EGFR) overexpression. Hwang et al. reported that E5 forms a complex with the EGFR and triggers the EGFR pathway activation nd generation of EGFR-TKI, is more pronounced in p16 negative tumors Decades ago, scholars initiated to block EGFR The heterogeneity of EGFR blockade therapy, in part, because of the downstream driver gene mutations, remains an incomplete understanding. PI3K mutation is highlighted in HPV-associated HNSCC. In the TCGA database, 27.8% PI3K mutated in HPV positive cases Aside from the PI3K pathway's stromal mutations, many studies reveal that E6 can target phosphorylase and activate the pathway. HPV E6 oncoprotein contains a PDZ-binding domain and inactivates PTEN Many pan-class I PI3K inhibitors have been developed and tested. The safety profile of inhibitors is acceptable Herein, finding a reasonable mode of combined therapy is the best choice for targeting PI3K. As the same as PI3K inhibitor, mTOR inhibitors often present a short-term efficacy It is well known that PI3K and MAPK pathways conduct in parallel. Combine PI3K with MAPK inhibitors in patients whose genetic alterations in both pathways coexist It has been reported that activation might be a prognostic marker for radiotherapy in HNSCC and indicates PI3K pathway inhibition would potentially exert a synergistic effect with radiotherapy Many trials have proved a worse clinical outcome in HPV-negative HNSCC. In RTOG 0129 trial, 433 patients with oropharyngeal cancer received cisplatin with radiotherapy. Compared with HPV-positive patients, the OS of HPV-positive tumor patients was remarkably worse . All the present data indicate that patients with HPV-negative tumors should be considered high-risk, and intensive, multimodal therapy is needed to avoid compromising their survival Genomic alterations are very common in HNSCC. HPV-negative cancers harbor significantly more mutations in the P53 gene than HPV-positive disease; meanwhile, loss-of-function variants in P53 are almost low to none in HPV-associated HNSCC As a transcription factor, P53 plays a crucial role in downstream target gene transcription. Under the stress of DNA damage, proto-oncogene activation, hypoxia, and microtubule damage, P53 is activated in the process of signal transduction. As a result, cellular behaviors, such as cell cycle arrest, apoptosis, aging, and angiogenesis inhibition, are performed to keep the benignancy of the cells'. P53 repairs the abnormal chromosome distribution in cells after DNA damage and mitosis in vivo and compromise tumor growth in the HNSCC animal model. INGN 201 is the first candidate of Ad-p53 for the Phase I clinical trial. The study recruited 33 patients with recurrent HNSCC and received INGN 201 intratumor injection. In the resectable cohort, 27% of HNSCC patients remained disease-free, with a median follow-up period of 18 months after surgery. Of the 17 unresectable diseases, in 2 cases had PR, 6 cases kept an SD, and 9 cases evaluated as PD. Multiple direct intratumor injection courses of INGN 201 were well tolerated, and no dose-limiting toxicity or serious adverse events were observed Decades ago, to overcome the loss function of mutated P53, people tried to reintroduce wild type P53 into the tumor via adenoviruses as vectors, and the virus can then selectively target and kill cancer cells. Adenovirus is safe and should be an ideal carrier for gene therapy as it can infect both mitoses and quiescent cells, and the genome keeps episomal and does not integrate into the host cells Loss-of-function P53 mutation is mainly located in the DNA binding domain, which effectively prevents the mutant p53 from binding to the target gene's response elements. p53-reactivating small molecules, including CP-31398 and PRIMA-1, are developed and tested in HNSCC Due to the PARP inhibitor's great success, scholars move their focus on hunting the synthetic lethal genes of P53. There is a complicated synthetic lethal relationship between mutated p53 and its corresponding target genes in all kinds of tumors. Based on the various research results of p53 synthetic lethality, the corresponding synthetic lethal relationship can be explained from two aspects: periodic regulatory genes and aperiodic regulatory genes It is well known that p53 activation can lead to cell cycle arrest and initiate DNA repair in response to DNA damage. Key protein inhibitors that regulate DNA damage response and cell cycle progression have the potential to be synthetic lethal partners ATM and ATR are members of the phosphatidylinositol 3-kinase-related kinase (PIKK) family of serine/threonine protein kinases. ATM and ATR are two critical factors to initiate the DNA damage response as a DNA damage sensor. ATM and ATR act as primary regulators of double-strand breaks and DNA replication stress, respectively. These two kinases perform functionally overlapping but non-redundant activities As major regulators of the ATM/ATR pathway, Chk1 and Chk2 kinases are functionally overlapping activated in response to DNA replication stress, cell cycle progression, chromatin remodeling, and apoptosis When DNA damage occurred in P53 wild type cells, the cell cycle checkpoints of G1 and G2/M are activated and prevents the accumulation of DNA damage. In P53 mutated cells, due to the dysfunction of the G/1 checkpoint, the G2/M cell cycle checkpoint is crucial when DNA is damaged Tumor-infiltrating lymphocytes are associated with improved prognosis. HNSCC arises from squamous epithelium associated with the tongue's tonsils and base and is deemed to have more immune cells infiltrated within the tumor microenvironment. HPV infection alters immune cell, which infiltrates in HNSCC to encompass diverse and heterogeneous landscapes HPV viral factors, E5, E6, and E7, play a crucial role in generating an immunosuppressive microenvironment that promotes tumor progression. Oncoprotein E5 blocks HLA-C and HLA-E from tumor stroma to interact with MHC I on the cancer cell, impairing T cell and NK cells' activity. Mandal et al. reported in an analysis of TCGA found that HPV-positive HNSCCs had more significant immune infiltration and had higher T cell levels with granzyme and perforin Promising clinical trial results led to pembrolizumab's FDA approval for treating refractory or metastatic HNSCC in 2016. Keynote 012 study was the first clinical trial to evaluate the relationship between HPV status and immune checkpoints blockade response. 60 patients with PD-L1+ (> 1%) were treated with pembrolizumab. ORR was 24%  in those patients with HPV positive, and 16% in HPV negative patients  In the phase III Checkmate 141 study, nivolumab obtained a better OS than standard treatment , with an approximately 19% increase for 1-year survival. In the Checkmate 141 subgroup analysis, patients with p16 positive tumors tend to obtain more benefit from nivolumab than the standard treatment group . On the other hand, in patients with p16 negative disease, the median OS was 7.5 months in the nivolumab monotherapy group and 5.8 months in the standard treatment group  However, the advantage of HPV status in nivolumab treatment failed to exhibit in pembrolizumab. In phase II KEYNOTE-055 trial, ORR was similar regardless of HPV status, with rates of 16% in HPV-associated disease and 15% in HPV-negative disease. However, medium PFS and OS did not differ based on HPV status Atezolizumab is an anti-PD-L1 mAb. In a phase I trial, 32 patients with R/M HNSCC were enrolled. The data were in line with anti-PD1 mAb. Atezolizumab monotherapy had a 22% ORR rate, mPFS of 2.6 months, and mOS of 6 months. However, this early trial's primary date displayed the effect of Atezolizumab, regardless of HPV status or PD-L1 expression level Although many retrospective or bio-informative studies indicated a higher degree of T cells in curium titration in HPV-associated HNSCC, the only nivolumab showed HPV+HNSCC could obtain more benefit from ICI therapy. The present clinical data indicate that HPV status does not support strategic treatment with checkpoint inhibitors.The imbalance, mutation, and aberrant expression of epigenetic regulatory factors boost carcinogenesis and maintain the growth and metastasis of HNSCC. Also, the levels of these epigenetic regulators varied with the type of HNSCC. From 2004, the US FDA approved only 6 epidrugs in the clinic: 2 DNMT inhibitors of azacitidine and decitabine; 4 HDAC inhibitors of Vorinostat, Romidepsin, Farydak, and Panobinostat. Many other agents are in the pipeline.Although scientists have done much work on the epigenetic regulation of HNSCC, rare epidrugs are in clinical trials. Many studies report that HPV-HNSCC has relatively low DNA methylation levels and promotes genomic instability. Meanwhile, HPV+ HNSCC harbors distinctly hypermethylated genomes. The diversity of molecular and epigenetic between HPV+ and HPV- tumors provides a therapeutic strategy that forces the demethylation of genomes of HPV associated NHSCC.DNMT inhibitors/DNA methylation agents present the cytotoxicity on HPV status bias. Asel Biktasova et al. reported that HPV+ HNSCC cells are sensitive to azacitidine partially due to stabilization of p53 and attenuation of the expression of HPV genes. Moreover, azacytidine is sufficient for suppressing cell invasion and sensitizing the cell to interferons. CUDC-101 is a small molecule that simultaneously blocks the EGFR, human growth factor receptor 2 (HER2), and histone deacetylase (HDAC) with promising activity in HNSCC. Thomas Galloway et al. reveals that CUDC-101 is tolerable in a phase I trial; however, a high discontinuation rate suggests CUDC-10 needs to alternate the schedules or routes of administration To sum up, we review the conditional therapy, including chemotherapy and radiotherapy, and indicate that HPV-positive patients may obtain more benefit from traditional treatment. More clinical trials need to be investigated for the optimal dosage for HPV-positive patients. PI3K is a promising target in HPV-positive HNSCC; however, the toxicity of combination therapy with PI3K inhibitor should not be ignored. Preclinical data present a suspected result of directly targeting P53, and synthetic lethality with DNA damage responder inhibitors remains too much to be explored. Immunotherapy is effective for HNSCC regardless of HPV status. At the present stage, as the same before 5 years, the HPV status can predict the prognosis more but cannot change the clinical decision-making. More prospective clinical trials are ongoing, which may answer critical questions over the next three to five years.


This is the most frequent sexually transmitted illness on the planet, and both men and women are equally vulnerable. HPV is associated with a broad variety of female disorders, including 99 percent of all cervical cancer cases. Specifically, the goal and contributions of this study are to determine Emirati men's opinions about the HPV vaccination, specifically whether they would use it themselves or allow their female relatives to use the vaccine. To collect the primary data, a statistical cross-sectional survey was conducted. This quantitative study was conducted using primary sources of data. A questionnaire survey with a sample size of 390 participants was used to collect data from 400 individuals. Male university students in the United Arab Emirati men have a weak grasp of HPV and are averse to vaccination . The percentage of Emirati men who accept the HPV vaccination is 37%. A total of 40.3% of the respondents opted not to participate in the survey at all. Eighty-six percent of the women surveyed had heard of cervical cancer, and one-third believed that they were at risk in the future. Twenty-five percent of those surveyed said that the HPV vaccination was safe, while 26% said it was unsafe. Respondents were just 3.1 percent vaccinated, and their family members were only 87% not vaccinated. Males and females alike are at risk for contracting this highly contagious illness. HPV is linked to a wide range of female diseases, including 99 percent of all cervical cancer cases, which is caused by HPV. Additionally, HPV is responsible for 60–90% of vaginal malignancies as well as 39% of vulvar cancers. Men's HPV infection is to blame for 39% of all penile cancers, while HPV has been linked to 39–65% of head and neck cancers and up to 88% of anal tumors in both sexes. Reduced HPV types including HPV 6 and 11 cause genital warts in both men and women, which have a substantial impact on quality of life and sometimes need extensive treatment. There is a strong correlation between HPV infection rates among affected males and their current sexual partners; thus, they are also at risk for additional high-risk HPV variations. Despite the lack of HPV research inside the Emirates (UAE), other Arabian Gulf nations have indicated a high prevalence of 11% for females [The FDA authorized the use of a trivalent HPV vaccination for men aged 9–26 years in October 2009. HPV vaccine for male genital warts and anal cancer has been authorized by the FDA and Advisory Committee on Immunization Practices, respectively. In addition, vaccination of men between the ages of 9 and 26 against HPV reduces the prevalence of genital warts, which has significant health and economic advantages. As a result, it has been claimed that male HPV vaccinations may protect their present or prospective spouses against cervical cancer as well as other HPV-related disorders [An HPV vaccination programmer for females entering grades 11 and 12 was introduced by Abu Dhabi's Health Authority (HAAD) in 2008. Inside the Middle East or Arab nations, Abu Dhabi is the very first state to implement HPV vaccination inside the public sector. An intensive media effort was also launched by Health Authority Abu Dhabi (HAAD), which arranged a training and awareness campaign for health care practitioners. The media and workplaces were used to spread the word. Previous research in the United Arab Emirates has shown that there is a significant lack of information regarding cervical screening and HPV vaccination among parents and healthcare professionals. Abu Dhabi's HPV vaccination rate started out at 50% but has now risen to 80%, according to HAAD. HAAD concluded that training medical professionals and educating women about the specifics of HPV infection were the most effective ways to increase HPV vaccine uptake. Studies have revealed that the most crucial element for good sentiments regarding HPV vaccination is actually knowing about it. The HPV vaccine has not yet been approved for boys in the United Arab Emirates; however, it is predicted that the health authorities in Abu Dhabi may consider providing the vaccine for guys following the completion of school immunization and a catch-up programmer. There are no studies to our knowledge that have looked at male HPV vaccination rates in the United Arab Emirates or even the Arabian Gulf states. The purpose of this research was to determine whether or not male Emirati men inside the United Arab Emirates are aware of and accept the HPV vaccine.There are approximately 1.9 billion Muslims in the world; with almost all Arab countries being Muslim, it has been always perceived that Muslims only practice sex through marriage as any extramarital relationship is forbidden; as such, the prevalence of any STI should be low and not warranting any preventative measures .Several research on human papillomavirus (HPV) inside the Middle East-North Africa area (MENA) region found that HPV prevalence was low compared to other western nations, until lately when certain studies showed an increase in the frequency of HPV. It was found that 17.8 percent of women in Oman had HPV infection  while anEstimates reveal that every year, 569,847 females are diagnosed with cervical cancer as 311,365 died from the condition. HPV infection is the most prevalent sexually transmitted infection in the world –15.The third most common malignancy in women worldwide is cervical cancer. Immediately after becoming sexually active, both men and women are at risk of contracting a sexually transmitted disease (STI). Sexual transmission of HPV does not need penetrative intercourse. Gnawing on each other's private parts is a well-known method of transmission as the most frequent HPV-related illness; cervical cancer is the most common. Cervical cancers are almost always caused by HPV infection, which accounts for the vast majority of these instances –18.Cervical cancer is caused by HPV in 99 percent of instances. Additionally, HPV is responsible for 60–90% of vaginal malignancies as well as 39% of vulvar tumors. Approximately to 39% of urethral malignancies are caused by HPV infection in men. HPV has been linked to 40–70% of head as well as neck cancers including up to 80% of anterior tumors in both sexes. Genital warts caused by low-risk HPV subtypes are common in both sexes and have a significant negative influence on one's quality of life. The sexual behavior in this hot region of the world is evolving rapidly in a risky manner, due to the introduction of new standards into this virgin community along with massive globalization, the wide spread use of information technology, migration, and political changes –21.There are several reasons why STI prevention is so important in the Arab world, which has one of the world's youngest populations. In 2008, Abu Dhabi's Health Authority for Abu Dhabi (HAAD) incorporated HPV vaccine in its immunization programmed. Vaccination for females entering grades 11 and 12 inside the Middle East or Arab nations, Abu Dhabi is first state to implement HPV vaccination inside the public sector .The present study is aimed at investigating the attitude of Emirati men towards HPV vaccine, whether they will use it themselves or allow their female relatives to use it.If you conceive of research as an umbrella phrase that incorporates a variety of tasks, you will be on the right track. In order to solve philosophical and practical questions, researchers use rigorous methods. According to the methodology and setting of the research, there are a variety of methodologies, including (a) descriptive analysis, which focuses on obtaining information that establishes the presence of what is being studied; (b) an evaluation and evaluation based on the review of information; (c) applicable analysis may be utilized in product, process, or policy development; and (d) foundational research is performed to fulfill scientific interest instead of for immediate practical application. There are many aspects that are not measurable and consequently cannot be estimated or interpreted quantitatively in qualitative research (e). There are much more instruments and procedures involved in quantitative research (f) than there are in qualitative research .In order to acquire the main data, a statistical cross-sectional survey was undertaken. Based on the original data, this statistical analysis was conducted. There were 390 participants in the questionnaire survey that collected data from 400 people.Analysis and interpretation of the acquired data were carried out using IBM SPSS. Descriptive research was used to effectively understand the study's findings  . ResearThe United Arab Emirates men constitute the study's research study population.The original data was used in this quantitative study; a sample of 390 survey respondents was employed to gather data, and a questionnaire survey was used to do so , 24.Prevalence of the taking HPV vaccine by respondents shows 37% acceptance rate which can be seen in HPV vaccine recommendation for respondents' daughters, relatives, or friends shows 46.7% yes response while 22.8% no response as given in We studied whether or not men in the United Arab Emirates were aware of and willing to get HPV vaccinations, as well as variables that enhanced or lowered their chance of doing so. Prevalence of the taking HPV vaccine by Emirati men shows 37% acceptance rate which also concise with the results of Mahmud et al. ; 40.3% oThe research revealed that males in the United Arab Emirates were aware of and willing to get HPV vaccines, as well as factors that increased or decreased their likelihood of doing so. The prevalence of HPV vaccination among Emirati males is 37%, which is consistent with Mahmud et al.'s  finding Data were obtained using a self-administered questionnaire and based on convenience sampling in our research. As a result, we decided to conduct a self-administered questionnaire in the conservative UAE community because of the cultural sensitivities of our questions. The low response rate of 71%, which is typical for self-administered surveys, might be a drawback.The study's key strength is that it is the first to examine sexual activities, understanding of HPV, and acceptance of HPV vaccination among UAE guys. There is a lack of data on sexual activity in our environment since sexual matters are often seen as taboo.For the research, it was determined whether or not males in the United Arab Emirates knew and were willing to acquire HPV vaccines and what factors influenced this decision. When it comes to getting the HPV vaccination, Emirati males had a 37% acceptance rate, which is in line with findings by Mahmud and colleagues , who fou


Ophiodon elongatus) is the subject of much speculation but little empirical research; ~20% of lingcod individuals exhibit this striking blue color morph, which is discrete from and found within the same populations as the more common brown morph. In other species, color polymorphisms are intimately linked with host–parasite interactions, which led us to ask whether blue coloration in lingcod might be associated with parasitism, either as cause or effect. To test how color and parasitism are related in this host species, we performed parasitological dissection of 89 lingcod individuals collected across more than 26 degrees of latitude from Alaska, Washington, and California, USA. We found that male lingcod carried 1.89 times more parasites if they were blue than if they were brown, whereas there was no difference in parasite burden between blue and brown female lingcod. Blue individuals of both sexes had lower hepatosomatic index  values than did brown individuals, indicating that blueness is associated with poor body condition. The immune systems of male vertebrates are typically less effective than those of females, due to the immunocompromising properties of male sex hormones; this might explain why blueness is associated with elevated parasite burdens in males but not in females. What remains to be determined is whether parasites induce physiological damage that produces blueness or if both blue coloration and parasite burden are driven by some unmeasured variable, such as starvation. Although our study cannot discriminate between these possibilities, our data suggest that the immune system could be involved in the blue color polymorphism–an exciting jumping-off point for future research to definitively identify the cause of lingcod blueness and a hint that immunocompetence and parasitism may play a role in lingcod population dynamics.The unusual blue color polymorphism of lingcod ( Biston betularia) shifted during the Industrial Revolution, such that dark-colored morphs that could camouflage against soot-blackened trees became favored over the light-colored morphs that had previously been dominant [In a textbook example of how the environment can shape expression of a color polymorphism, the selective landscape of the peppered moth  exhibits a dark orange coloration, which might arise because the trematode destroys the digestive tissues of the snail, releasing carotenoids that are deposited in the tissues of the foot [This example is not unique, as color and parasitism are intimately linked across animal taxa through a variety of mechanisms , 5; taxathe foot . Color pOphiodon elongatus) for whimsy . Despite the fact that ~20% of lingcod individuals on the west coast of North America exhibit this striking blue color morph , which is discrete from and found within the same populations as the more common brown morph , science has yet to explain why [Although color polymorphisms are common, few natural history oddities can match the blue color morph of lingcod  lingcod at the outset of this project, two of us noted that this species has among the highest average burdens of metazoan parasites that we have ever encountered in dissections of dozens of fish species . Furthermore, a parallel study uncovered differences in fatty acid signatures between blue and brown lingcod and found that blue lingcod were, on average, smaller than brown lingcod . BecauseNon-blue  lingcod might be more heavily burdened by parasites than blue lingcod for several reasons. If blueness in lingcod is an “honest signal” of resistance  against parasites (sensu ), then tAlternatively, blue lingcod might be more heavily burdened by parasites than brown lingcod. This could happen if blueness is an honest signal of tolerance to parasites. Tolerance refers to the ability of hosts to physiologically limit the fitness impacts of parasitism, and is distinct from resistance, or the ability to prevent parasitic infection in the first place . The andBlue lingcod might also carry heavier parasite burdens than brown individuals if blue coloration is a collateral impact of parasitism, resulting from the disruption of some organ system by infection. The liver or gallbladder are likely candidates, given that the biliverdin to which blue coloration is attributed is a bile pigment –15. In tSimilarly, behavioral differences among individuals might both mediate their exposure to agents that control coloration and their exposure to parasites. For example, some lingcod individuals might have dietary preferences for prey items that trigger biliverdin release or that have high levels of biliverdin in their tissues. If these prey items also carry high parasite burdens, then blue coloration and parasite burden could be positively correlated. In this case, we would only expect a difference between males and females in the frequency of the blue trait or in the parasite burden of blue versus brown individuals if one sex had a stronger dietary preference than the other for biliverdin-associated prey.Finally, perhaps body condition both induces the biliverdin release linked to blue coloration and simultaneously erodes immune defenses against parasitic infection . In otheGiven the rich variety of mechanisms that might link sex, coloration, and parasite burden, we were interested in assessing the associations among these variables. To that end, we sampled lingcod across more than 26 degrees of latitude from Alaska, Washington, and California, USA, and performed parasitological dissections on 89 lingcod individuals, representing a range of sizes, sexes, colors, depths of collection, and geographic locations.We sought to build on the findings of Galloway et al.  by creatFig 2). Note that samples for parasitological analysis were drawn from only four of these seven regions, spanning most of the geographic range: Southeast Alaska, Washington, northern California, and southern California. Three to four fishing ports were selected per region  based on location and accessibility of CPFVs. To ensure representation of lingcod across a wide range of size and age classes, we sampled shallow (<200 ft) and deep (200–550 ft) nearshore and offshore rocky reefs equally. On average, we fished for 2.5 days out of each port at different locations  to obtain the targeted sample size of 75–100 lingcod per port. Additional samples were provided by the Alaska commercial longline fishery, the NWFSC Rockfish Bycatch Study in Puget Sound [Lingcod  were collected between 2015 and 2017 using hook-and-line methods from chartered commercial passenger fishing vessels (CPFVs) from Yakutat, Alaska to San Diego, California, USA (54°30’N–34°30’N). The following seven regions were broadly identified: Southeast Alaska (54°30’N–59°48’N), Puget Sound and coastal Washington (46°16’N–49°N), Oregon (42°N–46°16’N), northern California (38°02’N–42°N), central California (34°30’N–38°02’N), and southern California , but color was noted as binary (brown or blue). If there was disagreement between internal and external coloration, we used the internal color, as it may represent a stronger phenotypic expression of the blue trait. To our knowledge, there is no scholarship on the process of post-mortem color degradation in lingcod, but we do know that internal blue coloration persists after death, because blue filets are sold in seafood markets and fetch a higher price than the more typical white filets; in fact, among seafood processors, it is known that blue coloration will remain in blue filets until cooking [We measured several phenotypic traits for each of the lingcod included in the coastwide dataset. Color was recorded as blue or brown immediately upon capture, then reassessed in the lab  . Color w cooking . For thiS1 Text). We identified all parasite taxa to the lowest possible taxonomic level using keys, host–parasite checklists, regional parasite identification guides, and primary literature.Within the coastwide group of lingcod, we sub-sampled 89 for parasitological study. These individuals were immediately measured and bagged upon capture to retain external parasites, which can otherwise drop off their host after death. For each of the 89 fish selected for parasitological dissection, we performed a comprehensive parasitological examination designed to detect most metazoan parasites. Processing time per fish ranged from 4 to 30 hours and averaged 16 hours. We did not search for myxozoan parasites, but all other metazoan parasites should have been detected by our standard parasitological dissection techniques , the color of the fish (brown or blue), and the interaction between sex and color. We included fish identification code nested within sampling location nested within sampling region  as a random intercept. We also included the interaction of sex * color with a random slope and random intercept varying across parasite taxa, to allow calculation of a unique estimate of the interaction effect for each parasite taxon. To account for the fact that larger fish carry more parasites than smaller fish, we included an offset term for log, which effectively converts the response variable to a rate . All numeric values were scaled prior to analysis by using the scale function in base R and then adding 2  and back-transformed after analysis to return them to their original units. We checked for collinearity by assessing whether variance inflation factors (VIF) were < 5 [glm.nb function in the lme4 package in R [We used generalized linear mixed models to assess the correlates of parasite burden for the subset of 89 lingcod selected for parasitological analysis. In these models, parasite abundance (number of parasites per fish) was the response variable. The generalized linear mixed-effects model took the form,were < 5 . Responsage in R .Fig 2), we sought to test whether the patterns revealed by the analysis above could have arisen due to random chance. We performed a permutation test in two iterations. In the first iteration, we randomly assigned four male fish in our dataset to be “blue” and the remainder to be “brown”. Females were not randomly assigned to a color, but instead retained their actual color assignment in the simulated dataset. This choice was made because the analysis above revealed extreme results for blue males and for no other combination of sex and color; we suspected that this might have been due to the exceptionally low number of blue males, and therefore tested the likelihood of obtaining such an extreme result with an identically sized random sample of males. After randomly appointing four “blue” males from among all the males in the dataset, we performed the analysis above and stored the resulting z-value for the interaction color * sex. We repeated this procedure 500 times and then calculated the proportion of runs in which the actual z-value for the interaction color * sex was more extreme (in either the positive or negative direction) than the simulated z-value for the interaction.Realizing that we had very low replication in one cell of our color * sex interaction  to be “blue” and the remainder to be “brown”. This choice was made because we surmised that the extreme results for blue males could have arisen by comparison to blue and brown females, and that random chance could have led to high rates of infection for brown females and low rates of infection for blue females, producing the significant interaction between sex and color. After randomly appointing four “blue” males from among all the males in the dataset and 11 “blue” females from among all the females in the dataset, we performed the analysis above and stored the resulting z-value for the interaction color * sex. We repeated this procedure 500 times and then calculated the proportion of runs in which the actual z-value for the interaction color * sex was more extreme (in either the positive or negative direction) than the simulated z-value for the interaction.ranef function in the lme4 package in R [Having investigated the overall patterns of parasite burden across parasite taxa, we then sought to understand whether individual parasite taxa differed in their responses to the interaction of host coloration and sex. From the model above, we extracted the random slope of the sex * color interaction for each parasite taxon and the variance associated with each slope, using the age in R . We thenadonis function in the vegan package in R [adonis does not support interaction terms), sampling location, and sampling depth.To assess the response of parasite community composition to host sex and color, we performed a multivariate analysis. We assessed whether parasite community composition varied among sex–color combinations with a PERMANOVA analysis, using the age in R . The PERijk represents either the Fulton’s K or hepatosomatic index of the ith fish collected from the jth site in the kth region. Fixed effects included: the depth of collection of the fish, the sex of the fish , the color of the fish (brown or blue), and the interaction between sex and color. We included sampling location nested within sampling region (to account for similarity among fish collected from the same sampling location or sampling region) as a random intercept. We excluded one outlier where an impossibly high body mass was accidentally recorded. All numeric predictors were scaled prior to analysis by using the scale function in base R and then adding 2  and back-transformed after analysis to return them to their original units. The model was run using the glmer function in the lme4 package in R [Finally, we performed two post-hoc tests. In light of our results (see below) and because Galloway et al.  detectedage in R .2 = 186.25, df = 1, p < 0.0001; Fig 3A). From among this initial group of lingcod collected, we analyzed the metazoan parasite burdens of 89 individuals, including 59 brown males, 4 blue males, 15 brown females, and 11 blue females. In these fish, we detected 29,465 parasite individuals across 25 taxa. A few parasite taxa dominated parasite detections. Specifically, larval trematodes (metacercariae) found in the fin and muscles of lingcod comprised 57% of all parasite detections. The next most abundant parasites were the adult trematode Rhipidocotyle elongata (9.8%), the larval acanthocephalan (cystacanth) Corynosoma wegeneri (5.8%), and copepod larvae . The remainder of the parasite taxa we detected (21 taxa) each comprised fewer than 5% of all parasite detections and included copepods, isopods, monogeneans, trematodes, cestodes, nematodes, and acanthocephalans (S1 Table). Although these 21 taxa comprised only a small proportion of total detections, these parasites still reached high abundances in infected hosts; for example, each of the 36 lingcod infected with cestode larvae (plerocercoids) in the order Trypanorhyncha carried on average 33 plerocercoids, and the lingcod with the heaviest burden carried 358 plerocercoids.We collected a total of 2,251 lingcod, of which 2,091 were definitively identified as either male or female and as either blue or brown. The blue trait was observed more frequently among females (27.2% blue) than among males . Averaging across blue and brown individuals within each sex, male lingcod had 1.88 times more parasites than females (Table 1), but this difference was driven almost entirely by blue males; brown males and brown females carried similar parasite burdens . Finally, lingcod collected from deeper depths tended to have lower parasite burdens than those collected from shallower depths (Table 1).Male lingcod carried 1.89 times more parasites if they were blue than if they were brown, whereas there was no difference in parasite burden between blue and brown female lingcod . In the first iteration of the permutation test, in which color was randomly permuted only among male lingcod, the probability of obtaining a result as or more extreme (in either direction) than the actual result was 3.4%. In the second iteration of the permutation test, in which color was randomly permuted among both male and female lingcod, the probability of obtaining a result as or more extreme (in either direction) than the actual result was 5%.Fig 4). In this analysis, a negative sex * color interaction indicates that the difference in parasite burden between blue and brown males is greater than the difference in parasite burden between blue and brown females; in other words, the more negative the interaction, the more parasites blue males carry compared to all other sex–color combinations. We extracted the random slopes for this interaction for each parasite taxon, and these are plotted in Fig 4, which shows that all parasite taxa were more abundant in blue males than in any other sex–color combination. Fin and muscle metacercariae had a weaker negative slope than any other parasite taxon, but that slope was still significantly different from zero, indicating that metacercariae are still more abundant in blue males than in any other sex–color combination, just to a lesser extent than other parasite taxa .The tendency for greater parasite abundance in blue males compared to all other sex–color combinations was consistent across all parasite taxa . Although PERMANOVA revealed no significant differences in community composition among sex–color combinations, we note that blue and brown males tended to differ in their burdens of fin and muscle metacercariae, Trypanorhyncha spp. plerocercoids, Corynosoma cetaceum cystacanths, Rhipidocotyle elongata adult trematodes, and chalimus copepod larvae .In alignment with the tendency for a consistent response to the sex * color interaction across parasite taxa, parasite community composition did not differ significantly among sex–color combinations, despite the higher total parasite burden in blue males . Our post-hoc analysis of the hepatosomatic index (HSI) revealed that, regardless of sex, blue lingcod had significantly lower HSI and hence reduced energy stores compared to brown lingcod . HSI increased with increasing depth (Table 2B).Finally, our post-hoc analysis of Fulton’s K revealed a marginally non-significant interaction between color and sex, with a trend for blue male lingcod to have lower Fulton’s K  than lingcod of any other sex–color combination . Below, we explore the elements of each mechanism that are supported (or not supported) by our data.Fig 1), so color is not confounded with geography, and blue males were collected across a broad subset (10.7–36.9 m) of the total depth range (7.6–72.2 m). These features of the data set imply that the pattern of high rates of parasitism among blue males is not an artifact of our sampling strategy and instead reflects a natural phenomenon.Blue males were encountered much less frequently than other sex–color combinations, resulting in a low sample size of that color morph. Despite this, the data suggest that blue male lingcod are characterized by heavier parasite burdens than other color–sex combinations in this species. The four highly parasitized blue males that drive this significant interaction were collected from geographically distant sites . Other examples of divergence between sexes in sensitivity to the pathological effects of parasitism exist; for instance, among honey bees (Apis mellifera) experimentally exposed to the gut parasite Nosema ceranae, males grow less and die sooner than females [Fig 4).Our data are also inconsistent with the hypothesis that blue coloration is an incidental outcome of parasitic infection, because we had originally assumed that parasitic infection would be equally physiologically disruptive for male and female lingcod, and that the sexes would therefore exhibit equal differences in burden between blue and brown individuals. However, if one sex  is likelier to die of parasitic infection, we would expect to see a smaller difference in burden between blue and brown individuals for the sex with the greater vulnerability. Alternatively, if the blue trait is initiated at different parasite thresholds between the sexes, we would expect to see a smaller difference in burden between blue and brown individuals for the sex with the lower threshold. If the parasite burden threshold that initiates blue coloration in females is very small, then it may not be detected in our study . On the other hand, perhaps males spend more time at depths or in habitats that expose them to both directly and trophically transmitted parasites [We cannot rule out the possibility that both blue coloration and parasite burden are driven by some unmeasured variable. In order to explain our observations, this variable would need to (1) make blue coloration more common in females and (2) drive a correlation between color and parasite burden in only males. Perhaps blue males are particularly likely to die of their heavy parasite burdens, making blue coloration appear more common in females. Alternatively, perhaps blue coloration is a reflection of diet differences between blue and brown lingcod  and thoarasites  and bluearasites , which marasites , 20. Finarasites , 20. NonFig 3C) suggests that low body condition is associated with blueness. We measured lower hepatosomatic index (HSI) values among blue fish than brown fish regardless of sex. These data suggest that low body condition  could cause blueness and, for male individuals whose immune systems are already compromised by male sex hormones, that low body condition may also increase susceptibility to parasitic infection–a conclusion that is supported by Galloway et al. [Our post-hoc analysis of lingcod body condition  suggests that whatever mechanism is at play affects most metazoan parasites equally. Although the response of fin and muscle metacercariae to the sex * color interaction was weaker than the response of the remainder of the parasites, it was nonetheless significantly negative, indicating that blue males carried a heavier burden of fin and muscle metacercariae than did brown males, blue females, and brown females. Therefore, whatever drives this effect acts in the same direction on all of the metazoan parasite taxa we detected. This strongly points to the immune system as a driver, and further supports the hypotheses above linking blue coloration with reduced immune efficacy in males.The fact that all parasite taxa contributed to the difference between blue and brown males  the effects we observed were consistent across all of the metazoan parasites we detected, and ecological drivers of the blue versus brown difference are unlikely to have consistent effects across such a great diversity of parasite taxa, and (2) effects were observable only in males, the sex that tends to have inherently lower immunocompetence across the vertebrates. It is possible that parasites induce physiological damage that produces blueness or that blue coloration and parasite burden may be driven by some shared physiological mechanisms, such as starvation. Although our study cannot discriminate among these possibilities, our data suggest the involvement of the immune system in the blue lingcod polymorphism–an exciting jumping-off point for future research to definitively identify the cause of lingcod blueness and a hint that immunocompetence and parasitism may play a role in lingcod population dynamics.S1 Fig(DOCX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.


Compared with the LBC test, the Cobas HPV test had higher sensitivity when detecting patients with high cervical intraepithelial neoplasia (CIN2+ and CIN3+).This study aimed to evaluate the performance of Cobas human papillomavirus (HPV) test in cervical cancer screening. A total of 3442 women aged ⩾20 years used Cobas HPV and hybrid capture 2 (HC2) tests were included in this study. Women with any positive result were examined by liquid-based cytology (LBC) test. Then subjects with abnormal LBC or positive Cobas HPV16/18 were further checked by colposcopy to observe the visible lesions to perform the pathological examination. Of these 3442 women, 328 cases were Cobas HPV positive, and the positive rate was 9.53%  8.50–10.53). The positive rate of HPV16, HPV18, and other 12 types of high-risk HPV were 1.54% (95% CI 1.12–1.95), 0.55% (95% CI 0.30–0.80), and 7.44% (95% CI 6.56–8.32), respectively. The coincidence rate of Cobas HPV test and HC2 test was 90%  in the primary screening. Age had a non-linear relationship with Cobas HPV positive rate ( Cervical cancer is the fourth most common cancer in women globally , with moin vitro for cervical cancer screening in recent years, which can detect 14 high-risk HPV subtypes, specifically report the results of high-risk HPV16 and 18 subtypes, and provide pooled results for the 12 other high-risk HPV subtypes  [Based on the pathogenicity, the high-risk HPV genotypes were divided into 15 types, which were closely related to cervical cancer and cervical intraepithelial neoplasia (CIN). They were HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68, respectively . The mai and 68) . In thisThis is a cross-sectional analysis, and a total of 7004 women were recruited into the Chinese public welfare project for two types of cancer screening  in Guangzhou Women and Children's Medical Center from December 2016 to June 2018. All of the participants were migrant workers, who lack regular cervical cancer screening. Before the HPV test, a questionnaire was conducted on the subjects’ education, marriage, contraceptive methods and cognition of cervical cancer. Participants were included if they met the following inclusion criteria: (1) subjects received both Cobas HPV test and HC2 test; (2) age ⩾20 years and (3) subjects with a history of sexual life, non-pregnancy, no history of cervical surgery. This study was approved by the Institutional Review Board (IRB) of the Guangzhou Women and Children's Medical Center, and written informed consent was obtained from all participants.Established files and registers, and provided informed consent and epidemiological investigations for women who met the screening conditions and voluntarily accepted the screening, and carried out Cobas HPV test and HC2 test (the primary screening). The positive results were detected by the liquid-based cytology (LBC) test (the second screening), and those with cytological abnormities or positive Cobas HPV16/18 were referred to colposcopy for examination. If necessary, a cervical biopsy was performed under colposcopy. The pathological diagnosis was performed by qualified pathologists in Guangzhou Women and Children's Medical Center. The accuracy of the screening methods was evaluated by pathologically confirmed cases of CIN2 or above.HC2 was a nucleic acid hybridisation detection method using microplate chemiluminescence for signal amplification, which could detect 13 types of high-risk HPV DNA  at one time by 96-well plate method, and simultaneously detect the viral load of HPV DNA in the samples. The HC2 test was performed by using the HC2 sample conversion kit (Qiagen) for the PreservCyt medium. If the relative light unit/cutoff ratio of the sample was ⩾1.0, the sample was recorded as positive.in-vitro quantitative detection technique for high-risk HPV DNA by polymerase chain reaction amplification, and can detect HPV16, 18 and other 12 types of high-risk HPV types , respectively. The Cobas HPV test was carried out according to the manufacturer's protocol [Cobas HPV test used the same specimens as the LBC test. Cobas HPV test is an protocol . InterprThe diagnostic criteria of the Bethesda System  , LBC andin situ. HSIL was characterised by thick acetowhite epithelium with clear boundary and was located near the junction of squamous column. Invasive carcinoma of cervix was featured with irregular, thick and brittle masses on the surface of the acetowhite epithelium. Cervical biopsy was performed under a colposcope. If lesions were visible under the colposcope, biopsy was taken directly at the lesion site, and if there were no visible lesions, biopsy was not performed.Five percent acetic acid solution was applied on the surface of the cervix. After 1 min, the changes in cervix were observed under a colposcope. Preliminary diagnosis was made according to the thickness, extent, surface morphology and turbidity of the acetowhite epithelium. Healthy cervix had no acetowhite epithelium. LSIL included HPV infection and CIN1, which has a light acetowhite epithelium and could be at or outside the junction of squamous column. HSIL included CIN2, CIN3 and cervical carcinoma t-test and expressed as mean ± s.d. Categorical variables were analysed by the chi-squared test (χ2 test) or the Fisher's exact test and displayed as number (n) and percentage (%). The Cohen's kappa was calculated to compare the HC2 test and Cobas HPV test results.SAS 9.4 software  was used for statistical analysis. Quantitative variables were tested by the P < 0.05 was considered statistically significant.When evaluating the screening efficacy of the Cobas HPV test and LBC test in detecting HSIL, pathological diagnosis was used as the gold standard and CIN2+ and CIN3+ were the disease endpoints respectively. MedCalc 14.8.1 software  was used to calculate the sensitivity, specificity, positive-predictive value, and negative-predictive value (NPV) of Cobas HPV test and LBC test. All statistical analyses were the two-sided test. The 95% confidence intervals (95% CIs) were calculated, and A total of 3442 women met the inclusion criteria . The meaP < 0.001) . The positive rate of HPV16, HPV18 and other 12 types of high-risk HPV were 1.54% (95% CI 1.12–1.95), 0.55% (95% CI 0.30–0.80) and 7.44% (95% CI 6.56–8.32), respectively. A total of 492 cases were found to be HC2 positive, and the positive rate was 14.29% (95% CI 13.09–15.51). The coincidence rate of Cobas HPV test and HC2 test was 90% (95% CI 89.00–91.00), which is medium-high consistency and statistically significant   and 3.Tχ2 = 4.240, P = 0.040). A ‘U-shaped’ distribution was shown in the relationship between age and Cobas HPV positive rate, and there were two peak age groups, 20–29 years old and 60+ years old respectively. There may be a non-linear relationship between age and HC2 positive rate, but the non-linear test was not statistically significant (P = 0.143)  .Fig. 2.χ2 = 6.610, P = 0.010). The age and HPV16/18 typing positive rate was a ‘U-shaped’ distribution, and there were two peak age groups, 20–29 years old and 60+ years old respectively. In addition, there may be a non-linear relationship between age and other 12 types of high-risk HPV typing positive rate, but the non-linear test was not statistically significant (P = 0.378)  .Fig. 3.Of these 582 positive cases in the first screening, 66 cases were also positive in the LBC test . The positive rate of ASC-US, LSIL, AGC, ASC-H and HSIL in the LBS test were 4.46% (95% CI 2.78–6.14), 1.89% (95% CI 0.78–3.00), 1.03% (95% CI 0.21–1.85), 2.03% (95% CI 0.88–3.18) and 1.72% (95% CI 0.66–2.78), respectively. The distribution of Cobas HPV positive rate in NILM, ASC-US, LSIL, AGC, ASC-H and HSIL were 46.38% (95% CI 42.33–50.43), 3.96% (95% CI 2.38–5.54), 1.89% (95% CI 0.78–3.00), 0.51% (95% CI 0.0–1.09), 2.06% (95% CI 0.91–3.21) and 1.52% (95% CI 0.53–2.51), respectively. The detailed results were shown in Among 54 positive cases after two screenings, 20 cases were positive  according to the pathological diagnosis. The incidence rates of CIN1, CIN2, CIN3 and squamous/adenocarcinoma were 5.56% (95% CI 0.0–11.73), 12.96% (95% CI 4.00–21.92), 11.12% (95% CI 2.73–19.51) and 7.40% (95% CI 0.42–14.38), respectively .Table 6vs. , P = 0.002) and NPV  vs. , P = 0.042) were higher in the Cobas HPV test than those in the LBC test. Similarly, when CIN3+ is used as the disease endpoint, compared with the LBC test, the sensitivity  vs. , P = 0.009) and NPV  vs. , P = 0.315) were also higher in the Cobas HPV test. The Cobas HPV test is more effective at detecting CIN2+ and above than the LBC test  LBC test  and 8.TIn this study, we evaluated the performance of Cobas HPV test for cervical cancer screening in female migrant workers. The positive rate detected by the Cobas HPV test was 9.53%, and the coincidence rate of the Cobas HPV test and HC2 test was 90% in the primary screening. The Cobas HPV positive rate and HPV16/18 typing positive rate had a non-linear relationship with age. A ‘U-shaped’ distribution was shown in the relationship between age and Cobas HPV positive rate and HPV16/18 typing positive rate, and there were two peak age groups, 20–29 years old and 60+ years old respectively. Compared with the LBC test, the Cobas HPV test had higher sensitivity and NPV when detecting patients with CIN2+ and CIN3+.Persistent high-risk HPV infection is one of the leading causes of cervical carcinoma and related deaths in women worldwide. It was reported that HPV16 and HPV18 in high-risk HPV types could cause about 70% of cervical squamous cell carcinomas and about 80% of cervical adenocarcinomas . In Chinet al. showed that the best outcomes for detection of disease were seen using the Cobas HPV test compared with the LBC test [Our study also compared the performance of Cobas HPV test and LBC test in detecting HSIL. The Cobas HPV test had higher sensitivity and NPV than the LBC test when detecting CIN2+ and CIN3+ patients. A prospective study performed that the Cobas HPV test had higher sensitivity than the LBC test in detecting CIN2+ and CIN3+ lesions, and the combination use of Cobas HPV test and LBC test can improve specificity . Dreyer LBC test .In this study, most women who participated in cervical cancer screening were married women who made a living by labour work in the city. Most of them were in the service industry and lacked the awareness of active physical examinations . In addiThe Cobas HPV test was more sensitive than LBC in the application of cervical cancer screening, especially in a large-scale population with a high potential infection rate. Compared with the HC2 test, Cobas HPV reported the results of HPV16 and 18 while detecting the high-risk HPV, which is helpful for doctors to refer high-risk groups to colposcopy in time, and to detect and treat patients with high risks of pathological changes earlier.


Low-grade cervical squamous intraepithelial lesion is a precancerous neoplasia that has appreciable probability to evolve into malignancy. To explore the prognostic value of HPV 16/18 genotyping and geminin mRNA quantification in predicting the progressiveness of LSIL. We recruited 212 participants who were negative for intraepithelial lesion or malignancy (NILM 76), low-grade squamous intraepithelial lesion (LSIL 85), high-grade squamous intraepithelial lesion (HSIL 36) and cervical intraepithelial neoplasia grade cervical cancer grade 3, (CIN3 15) patients. Tissues were obtained during excisional treatment. HPV 16/18 genotyping and geminin mRNA qRT-PCR were performed. HPV 16/18 positivity rate and geminin mRNA level were integrated with the clinical parameters into a multivariate logistic model. Area under curve was yielded based on receiver operation curve derived from this multivariate logistic model. Follow-up visits were performed to LSIL patients with progression. HSIL patients have higher HPV 16/18 positivity rate and geminin mRNA levels than LSIL. Among HSIL, CIN3 have higher HPV 16/18 positivity rate and geminin mRNA levels. Multivariate logistic analysis showed that HPV 16/18 positivity and geminin mRNA expression status are independent factors for differentiating HSIL and LSIL. The baseline HPV 16/18 positivity rate and geminin mRNA levels of 18 LSIL patients who developed HSIL are significantly higher than non-progressive LSIL patients. The values examined at follow-up timepoints were also higher than baseline. These results suggest that geminin is implicated in the progression of LSIL and combining HPV 16/18 genotyping and geminin mRNA qRT-PCR could potentially differentiating the progressive LSIL and improve the efficacy of clinical intervention. Cervical cancer is a common malignancy that originates in women’s reproduction system, which posts a threat on women’s life and well-being . Recent Currently, cytology is the most reliable method to predict progression of LSIL. Normal cytology ) or mildly abnormal cytology, i.e. atypical squamous cells of undermined significance has relatively low 5-year progression rate, while more severe cytological abnormalities, i.e. atypical squamous cells or atypical glandular cells, holds substantially higher progressive risks. However, 10% progression was developed from LSIL with normal or mildly abnormal cytology . In clinHuman papilloma virus (HPV) is the most common sexually transmitted infection in the world, which is also the most prevalent cause of cervical carcinoma and CIN . In recePatients with LSIL, HSIL or cervical cancer who were admitted to Jiangxi Cancer Hospital from December 2016 to November 2019 were recruited for this study.Women with HSIL, LSIL/CIN1 had been diagnosed by colposcopy-guided biopsy. The inclusion criteria: (1) Within 3 days, no sexual intercourse was conducted, no vaginadouche or administration was performed; (2) Nohistory of CIN diagnosis; (3) Normal liver and kidney function. Exclusion criteria: (1) complication of other morbidities such as diabetes, chronic kidney disease, and heart failure; (2) administrated with immune inhibitor drugs; (3) women in pregnancy or lactation period. In total, we recruited 212 participants, in which 76 are negative for intraepithelial lesion or malignancy, 85 were diagnosed with LSIL, 36 were HSIL, and 15 cervical cancer patients. In LSIL patient group, the mean age was 38.36 ± 7.32, ranging between 27 and 65 years; the average childbearing was 2.14 ± 0.32 times, and delivery 1.39 ± 0.21 times; in HSIL patient group, the mean age was 39.46 ± 6.83, ranging between 26 and 68 years; the average childbearing was 2.64 ± 0.42, and delivery 1.62 ± 0.25.Cervical tissues were obtained during the excisional treatment and were fixed in 10% neutral buffered formalin and embedded in paraffin following routine procedures. This study was approved by the Ethic Committee of Jiangxi Cancer HospitalHospital, and informed consent were obtained from all participants.For HPV genotyping, DNA was extracted from the stored SurePath Pap specimens using the DNeasy kit  according to the manufacturer’s instructions. HPV genotyping was conducted using the the TRUPCR® HPV High Risk (E6/E7) Kit, which is an RT-qPCR assay based on oligonucleotide hydrolysis principle that allows higher specificity and sensitivity of E6/E7 region by primer and probes specific for HPV 14 Genotypes (16/18/31/33/35/39/45/51/52/56/58/59/66 and 68) and simultaneous genotyping of 16 and 18.Complementary DNA was synthesized from total RNA using a M-MLV reverse transcription kit  with the specific primers of GMNN. cDNA samples were used as templates for amplification reactions performed in a PCR Thermal Cycler Dice Real-Time system with the SYBR PrimeScript PCR kit (Takara Bio).All qPCRs were performed in triplicate at a reaction volume of 25 µL containing 5 µL of cDNA, diluted at a 1:5 ratio, and mixed with Taqman Universal PCR MasterMix. The following protocol was used for all assays: denaturation (95°C for 10 min) and amplification  repeated for 40 cycles. The housekeeping genes GUSB (beta glucuronidase) and PGK1 (cGMP-dependent protein kinase (1) were selected as reference genes for quality control of the RNA specimens. This combination of reference genes demonstrated a high stability in expression between groups of normal samples versus HSIL samples. The number of cycles required for the signal to cross the threshold  for target genes was set at 35 cycles and automatically calculated and recorded by the High-Resolution Melt Software v2.0 for each reaction. Ct levels are inversely proportional to the amount of target nucleic acid in the sample . For the reference genes GUSB and PGK1, a Ct value above 35 cycles indicates poor RNA quality. Expression of GMNN were analyzed with 2−ΔΔCt method. Each sample was quantified in triplicate, and mean values were adopted for further analysis. Expression of GMNN were normalized to endogenous GPDH control.t-test. Enumeration data was represented with frequency (%) and analyzed with Chi-square nonparametric test. Variables that were significant in univariate analysis were further testified in multivariate logistic regression. Receiver Operating Characteristic Curve (ROC) was employed to verify the diagnostic effectiveness of our logistic prediction model, from which Area Under Curve (AUC) and cutoff values were derived. P < 0.05 was considered statistical significant.SPSS 17.0 software was adopted to perform statistical analysis for the data. Normally distributed data were presented as Mean ± SD. Group comparison was analyzed with Student’s The basic aim of the study was to explore the prognostic value of HPV 16/18 genotyping and geminin mRNA quantification in predicting the progressiveness of LSIL. We recruited 212 participants who were negative for intraepithelial lesion or malignancy (NILM 76), Low-grade squamous intraepithelial lesion (LSIL 85), high-grade squamous intraepithelial lesion (HSIL 36) and Cervical intraepithelial neoplasia grade cervical cancer grade 3, (CIN3 15) patients. Tissues were obtained during excisional treatment. HPV 16/18 genotyping and geminin mRNA qRT-PCR were performed. HPV 16/18 positivity rate and geminin mRNA level were integrated with the clinical parameters into a multivariate logistic model. Area under curve was yielded based on receiver operation curve derived from this multivariate logistic model. Follow-up visits were performed to LSIL patients with progression.HPV 16/18 mRNA cannot be detected in normal cervical tissues, but was expressed in LSIL, HSIL and cervical cancer tissues . The posp = 0.003; LSIL vs. cervical cancer, p = 0.002). HSIL/CIN3 expressed higher geminin than HSIL/CIN2 (p = 0.001).LSIL tissues were detected with low level of geminin, and both HSIL and cervical cancer tissues demonstrated substantially higher geminin mRNA expression  is visible in most of NILM and LSIL participants, but only present in 22% (8/36) of HSIL participants and not even visible in all cervical cancer patients. Overall, HSIL and cervical cancer patients had larger proportion of medium/large size lesions as per TNM classification. A total of 64.7% LSIL patients have small lesion, 24.7% have medium size lesion, and 10.6% have large size lesion. In HSIL group, 13.9% have small lesion, 41.7% have medium size lesion, and 44.4% have large lesion. In addition to the substantial difference of HPV 16/18 positivity mentioned above, high geminin level was found in 30.6% LSIL participants, 80.6% HSIL and 93.3% cervical cancer patients.Variables that demonstrated significant difference between the disease groups have been assessed by the multivariate logistic regression model. Results showed that HPV 16/18 positivity status, geminin expression, SCJ presence and lesion size are independent predictor that differentiating LSIL and HSIL, in which HPV 16/18 status and geminin rank as top 2 factors . We nextGiven that HPV 16/18 and geminin hold potential as a prognostic and diagnostic biomarker for differentiating HSIL from LSIL, whether HPV 16/18 and geminin expression change during the progression of LSIL/CIN1 remains to be discovered. Follow-up was done at 6-month intervals for up to 2 years. At each follow-up, women positive for cytology or high-risk HPV (hrHPV) were referred for colposcopy To address this, we followed up 18 women who had LSIL but progressed to HSIL/CIN2 or HSIL/CIN3. Cytology and expression of HPV 16/18 were evaluated. HPV 16/18 positivity and geminin levels were significantly increased as compared with their baseline expression measured . The nonDifferentiation of LSIL and HSIL can be challenged because the lesion of them are histologically continuous and identifying potential progressive LSIL is even more complicated and requires close follow-up when the progression can be detected. The widely use of LEEP could eradicate the progressive LSIL which further may increase the economic burden to individual, family, and society because of its limited affordability. Therefore, seeking for differentiating biomarker is of great importance for reducing incidence of cervical cancer.HPVis a heterogeneous group of viruses that are common in human world that comprises of more than 100 types. Among them, 14 types of HPV were found related to cancer, such as cancer of the anus, vulva, vagina, penis and oropharynx . CervicaGeminin plays an essential role in embryonic development and maintenance of chromosomal integrity. Although it was reported as a protective gene from oncogenesis, multiple lines of evidence demonstrated geminin overexpression in human cancers, suggesting its ambivalent role in cancers. A recent study demonstrated that geminin is aberrantly overexpressed in breast cancer tissues and promotes tumor invasionand metastasis by suppressing FoxO3-mediated transactivation of Dicer . Kim et We found that HPV 16/18 positivity and geminin level are two independent factors with the most significant OR. The detective effectiveness of this model gauged by the area under curve (AUC) of the receiver operation curve (ROC) yielded an accuracy of 0.932 with sensitivity of 0.912 and specificity of 0.897. Taken together, our study explored the prognostic value of combining HPV 16/18 genotyping and geminin mRNA quantification in differentiation of progressive LSIL, which lends real-world evidence for developing diagnostic tool for LSIL.