Normal Cervix Research Articles

Evaluation of Automatic Signal Detection of In Situ Hybridization for Detecting HPV DNA in Cervical Tissue Derived from Patients with Cervical Intraepithelial Neoplasia

Background: Cervical cancer is fourth the most common cancer in women worldwide. Due to the prevalence of human papillomavirus (HPV) in the population (80–90%), scientists are likely to discover even more associations of this pathogen with other diseases in the future. In recent years, In Situ Hybridization (ISH) assays that use automated signal-detecting methods in formalin-fixed, paraffin-embedded (FFPE) cervical tissue, such as the enzyme-categorized signal-detecting system, have shown a higher sensitivity. Objectives and Methods: To evaluate automatic signal detection of ISH assay for detecting HPV DNA, we compared the ability of an ISH probe, Inform HPV II and III (Ventana Medical Systems, Tucson, AZ), to that of PCR assays to detect HPV DNA in cervical tissue specimens with cervical intraepithelial neoplasia (CIN; CIN 1, 28 cases; CIN 2, 22 cases; and CIN 3, 20 cases) and normal cervix (2 cases). Results: Our findings showed a significant relation was confirmed between ISH III level and HPV outcome (positive/negative). Patients with positive HPV outcomes had significantly lower ISH III levels, MD = −7961.82 CI95 [−17,230.00; −199.21], p = 0.005. Conclusions: Automatic signal detection of ISH assay is not particularly applicable to cervical tissue material. A more useful method of confirming the presence of HPV in the cervix is the HPV test with genotyping, as it allows for collecting a larger amount of material from the cervical disc and canal. The interpretation of a positive or negative ISH test must be guided in the context of clinical history and morphology.

The Correlation of Hysteroscopy and Histopathology in Evaluating Abnormal Uterine Bleeding: Observations From a Tertiary Care Center.

Abnormal uterine bleeding (AUB) is an important clinical entity, and its subtype, heavy menstrual bleeding (HMB), affects 14%-25% of women of reproductive age, potentially impairing their physical, emotional, social, and material quality of life. However, in addition to prior studies that supported the efficacy of hysteroscopy in identifying AUB, this study determined the overall pattern of abnormalities in AUB and correlated the diagnosis through hysteroscopy and histopathology. Additionally, a comparison of ultrasonography was done in this study. This study helps healthcare providers provide timely diagnosis and, thereby, timely interventions in treating different causes of AUB. This hospital-based prospective observational study was conducted at a tertiary care center, including women aged 20-60 admitted with complaints of AUBbetween September 2022 and June 2024. Participants were selected based on their willingness to participate in the study. A statistical analysis was performed using relevant descriptive statistics andplotting the frequency and percentage. Cohen's kappa was applied to ascertain significant associations and patterns within the dataset. Results: The study included 47 women who were admitted to the hospital for diagnostic hysteroscopy. A majority of clinical symptoms presented as menorrhagia (17, 36.2%). The hemoglobin (g/dL) ranged from 5.9 to 14.7. Overall, on hysteroscopy, 37 (78.7%) women had a normal uterus. On hysteroscopy, 36 (76.6%) participants showed a normal cervical canal and cervix. In terms of histopathology, a majority of the study population (21, 44.7%) had proliferative endometrium. In terms of the correlation between hysteroscopy and histopathology, the two methods agreed in 83% of cases and disagreed in 17% of cases. Thisstudy confirms that hysteroscopy is a crucial tool for assessing patients with AUB, particularly those with a thick endometrium, across all age groups. It does not replace other diagnostic methods; instead, it enhances them. Hysteroscopic-guided biopsy along withhistopathology is now widely regarded as the most reliablemethod and new gold standardfor assessing cases of AUB.

An analysis of the vaginal microbiota and cervicovaginal metabolomics in cervical lesions and cervical carcinoma

BackgroundTo explore the role of vaginal microbiota and metabolomics in the progression of cervical dysplasia. MethodsThe patient group consists of female patients with low-grade, high-grade cervical dysplasia, and cervical cancer. Normal cervix samples from health volunteers were used as controls. The metabolic fingerprints of cervicovaginal lavage were analyzed using liquid chromatography-mass spectrometry, while the vaginal microbiota was examined through 16S rRNA sequencing. Bioinformatic analysis was adopted to investigate the interplay between hosts and microbes. The vaginal metabolic and microbiota profiles of 90 female patients with cervical dysplasia and 10 controls were analyzed to discover the biological characteristics underlying the progression of cervical cancer. ResultsWe found that Valyl-Glutamate, N, N′-Diacetylbenzidine, and Oxidized glutathione, which were involved in oxidative stress response, were discriminators to distinguish the normal cervix, invasive cervical carcinomas, and CIN3 from others. Cervical carcinoma was characterized by a large variety of vaginal microbes (dominated by non-Lactobacillus communities) compared to the control. These microbes affected amino acid and nucleotide metabolism, producing metabolites with cervical carcinoma and genital inflammation compared to the control group. ConclusionsThis study revealed that cervicovaginal metabolic profiles were determined by cervical cancer, vaginal microbiota, and their interplays. ROS metabolism can be used to discriminate normal cervix, CIN3, and invasive cervical carcinoma.

TPP1 is associated with risk of advanced precursors and cervical cancer survival.

It is unclear how telomere-binding protein TPP1 interacts with human telomerase reverse transcriptase (hTERT) and influences cervical cancer development and progression. This study included all eligible 156 cervical cancers diagnosed during 2003-2008 and followed up through 2014, 102 cervical intraepithelial neoplasia (CIN) patients, and 16 participants with normal cervix identified at the same period. Correlation of expression of TPP1 and hTERT in these lesions was assessed using Kappa statistics. TPP1 was knocked down by siRNA in three cervical cancer cell lines. We assessed mRNA expression using quantitative real-time polymerase chain reaction and protein expression using tissue microarray-based immunohistochemical staining. We further analyzed the impact of TPP1 expression on the overall survival of cervical cancer patients by calculating the hazard ratio (HR) with 95% confidence intervals (CIs) using the multivariable-adjusted Cox regression model. Compared to the normal cervix, high TPP1expression was significantly associated with CIN 3 and cervical cancers (P<0.001 for both). Expressions of TPP1 and hTERT were highly correlated in CIN 3 (Kappa statistics = 0.50, P = 0.005), squamous cell carcinoma (Kappa statistics = 0.22, P = 0.011), and adenocarcinoma/adenosquamous carcinoma (Kappa statistics = 0.77, P = 0.001). Mechanistically, knockdown of TPP1 inhibited the expression of hTERT in both mRNA and protein levels. High expression of TPP1 (HR = 2.61, 95% CI 1.23-5.51) and co-high expression of TPP1 and hTERT (HR = 2.38, 95% CI 1.28-4.43) were independently associated with worse survival in cervical cancer patients. TPP1 and hTERT expression was correlated and high expression of TPP1 was associated with high risk of CIN 3 and cervical cancer and could predict a worse survival in cervical cancer.

3D MR elastography-based stiffness as a marker for predicting tumor grade and subtype in cervical cancer

BackgroundIncreasing evidence has indicated that high tissue stiffness (TS) may be a potential biomarker for evaluation of tumor aggressiveness. PurposeTo investigate the value of magnetic resonance elastography (MRE)-based quantitative parameters preoperatively predicting the tumor grade and subtype of cervical cancer (CC). Study typeRetrospective. PopulationTwenty-five histopathology-proven CC patients and 7 healthy participants. Field strength/sequence3.0T, magnetic resonance imaging (MRI) (LAVA-flex) and MRE with a three-dimensional spin-echo echo-planar imaging. AssessmentThe regions of interest (ROIs) were manually drawn by two observers in tumors to measure mean TS, storage modulus (G′), loss modulus (G″) and damping ratio (DR) values. Surgical specimens were evaluated for tumor grades and subtypes. Statistical testsIntraclass correlation coefficient (ICC) was expressed in terms of inter-observer agreements. t-test or Mann-Whitney nonparametric test was used to compare the complex modulus and apparent diffusion coefficient (ADC) values between different tumor groups. Area under the receiver operating characteristic curve (AUC) analysis was used to evaluate the diagnostic performance. ResultsThe TS of endocervical adenocarcinoma (ECA) group was significantly higher than that in squamous cell carcinoma (SCC) group (5.27 kPa vs. 3.44 kPa, P = 0.042). The TS also showed significant difference between poorly and well/moderately differentiated CC (5.21 kPa vs. 3.47 kPa, P = 0.038), CC patients and healthy participants (4.18 kPa vs. 1.99 kPa, P < 0.001). The cutoff value of TS to discriminate ECA from SCC was 4.10 kPa (AUC: 0.80), while it was 4.42 kPa to discriminate poorly from well/moderately differentiated CC (AUC: 0.83), and 2.25 kPa to distinguish normal cervix from CC (AUC: 0.88), respectively. There were no significant difference in G″, DR and ADC values between any subgroups except for comparison of healthy participants and CC patients (P = 0.001, P = 0.004, P < 0.001, respectively). Data conclusion3D MRE-assessed TS shows promise as a potential biomarker to preoperatively assess tumor grade and subtype of CC.

Single-cell RNA sequencing of cervical exfoliated cells reveals potential biomarkers and cellular pathogenesis in cervical carcinogenesis

Cervical cancer (CC) is a common gynecological malignancy. Despite the current screening methods have been proved effectively and significantly decreased CC morbidity and mortality, deficiencies still exist. Single-cell RNA sequencing (scRNA-seq) approach can identify the complex and rare cell populations at single-cell resolution. By scRNA-seq, the heterogeneity of tumor microenvironment across cervical carcinogenesis has been mapped and described. Whether these alterations could be detected and applied to CC screening is unclear. Herein, we performed scRNA-seq of 56,173 cervical exfoliated cells from 15 samples, including normal cervix, low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL), and malignancy. The present study delineated the alteration of immune and epithelial cells derived during the cervical lesion progression. A subset of lipid-associated macrophage was identified as a tumor-promoting element and could serve as a biomarker for predicting the progression of LSIL into HSIL, which was then verified by immunofluorescence. Furthermore, cell–cell communication analysis indicated the SPP1-CD44 axis might exhibit a protumor interaction between epithelial cell and macrophage. In this study, we investigated the cervical multicellular ecosystem in cervical carcinogenesis and identified potential biomarkers for early detection.

The role of cervical elastography in the differential diagnosis of preinvasive and invasive lesions of the cervix.

This study aims to evaluate the role of cervical elastography in the differential diagnosis of preinvasive and invasive lesions of the cervix. A total of 95 women participated in this prospective study and were divided into the following groups: 19 healthy subjects (group 1) with normal cervicovaginal smear (CVS) and negative human papillomavirus test (HPV DNA), 19 women with normal cervical biopsy and normal final pathological result of cervical biopsy (group 2), 19 women with low-grade squamous intraepithelial lesion (LSIL) (group 3), 19 women with high-grade squamous intraepithelial lesion (HSIL) (group 4), and 19 women with cervical cancer (group 5). Clinical, demographic, histopathological, and elastographic results were compared between these groups. Comparing groups, age (40.42 ± 8.31 vs. 39.53 ± 8.96 vs. 38.79 ± 9.53 vs. 40.74 ± 7.42 vs. 54.63 ± 12.93, p < 0.001 respectively), gravida (1.74 ± 1.33 vs. 2.16 ± 1.68 vs. 2.21 ± 1.96 vs. 2.53 ± 1.93 vs. 4.63 ± 2.17 p < 0.001 respectively), parity (1.37 ± 0.68 vs. 1.68 ± 1.20 vs. 1.58 ± 1.30 vs. 2.00 ± 1.67 vs. 3.37 ± 1.61, p < 0.001 respectively), and the proportion of patients at menopause (10.5% vs., 15.8% vs. 10.5% vs. 5.3% vs. 57.9%, p < 0.01 respectively), a statistically significant difference was found (Table1). However, no statistically significant difference was found in the number of abortions, BMI, mode of delivery, smoking, additional disease status, history of surgery, and family history (p > 0.05) (Table2. As a result of the applied roc analysis, mean cervical elastographic stiffness degree (ESD) was found to be an influential factor in predicting cervical cancer (p < 0.05). The mean cut-off value was 44.65%, with a sensitivity of 94.7% and a specificity of 96.1% (Table7). Measurement of ESD by elastography is a low-cost, easily applicable, and non-invasive indicator that can distinguish cervical cancer from normal cervical and preinvasive lesions. However, it is unsuitable for determining preinvasive cervical lesions from normal cervix.

Labeled-free quantitative proteomic analysis of cervical squamous cell carcinoma identifies potential protein biomarkers.

Cervical cancer remains a prevalent cancer among women, and reliance on surgical and radio-chemical therapies can irreversibly affect patients' life span and quality of life. Thus, early diagnosis and further exploration into the pathogenesis of cervical cancer are crucial. Mass spectrometry technology is widely applied in clinical practice and can be used to further investigate the protein alterations during the onset of cervical cancer. Employing labeled-free quantitative proteomics technology and bioinformatics tools, we analyzed and compared the differential protein expression profiles between normal cervical squamous cell tissues and cervical squamous cell cancer tissues. GEPIA is an online website for analyzing the RNA sequencing expression data of tumor and normal tissue data from the TCGA and the GTEx databases. This approach aided in identifying qualitative and quantitative changes in key proteins related to the progression of cervical cancer. Compared to normal samples, a total of 562 differentially expressed proteins were identified in cervical cancer samples, including 340 up-regulated and 222 down-regulated proteins. Gene ontology functional annotation, and KEGG pathway, and enrichment analysis revealed that the differentially expressed proteins mainly participated in metabolic pathways, spliceosomes, regulation of the actin cytoskeleton, and focal adhesion signaling pathways. Specifically, desmoplakin (DSP), protein phosphatase 1, regulatory (inhibitor) subunit 13 like (PPP1R13L) and ANXA8 may be involved in cervical tumorigenesis by inhibiting apoptotic signal transmission. Moreover, we used GEPIA database to validate the expression of DSP, PPP1R13L and ANXA8 in human cancers and normal cervix. In this study, we identified 562 differentially expressed proteins, and there were three proteins expressed higher in the cervical cancer tissues. The functions and signaling pathways of these differentially expressed proteins lay a theoretical foundation for elucidating the molecular mechanisms of cervical cancer.

Correlations between C-myc expression, BMI-1 expression, and vaginal microecology with HPV-DNA load in patients with different cervical lesions.

To investigate the correlations between the expressions of proto-oncogenes C-myc and B-cell-specific Moloney leukemia virus integration site-1 (BMI-1), vaginal microecology, and human papillomavirus-DNA (HPV-DNA) load in patients with different cervical lesions. A total of 51 patients with cervix squamous cell carcinoma (CSCC), 72 patients with cervical intraepithelial neoplasia (CIN) and 50 patients with normal cervix (NC) who were diagnosed or admitted between Jan. 1st 2020 and Dec. 31st 2022 at the Suzhou Hospital of Integrated Traditional Chinese and Western Medicine were selected and divided into three groups, i.e., the CSCC group, the CIN group and the NC group, for a retrospective analysis. Hybrid capture 2 (hc2) was used to detect the HPV-DNA load in each group. Immunohistochemistry was performed to detect C-myc and BMI-1 expressions in each group. The indicators of vaginal microecology in patients were compared among groups to analyze the correlations between C-myc, BMI-1 expressions, vaginal microecology and HPV-DNA load. The HPV-DNA load and expression levels of positive C-myc and BMI-1 in the CSCC group were all higher than those of the CIN and NC groups (P<0.05). The detection rate of lactobacillus in the CSCC group was lower than that of the CIN and NC groups. The percentages of leukocyte esterase (LE) positivity and pH ≥4.6 were higher in the CSCC group than those in the CIN and NC groups (P<0.05). The difference in the detection rate of spores among the three groups was not significant (P>0.05). Both C-myc and BMI-1 scores were positively correlated with HPV-DNA load in the 173 samples. The proto-oncogenes C-myc and BMI-1 were highly expressed in the cervical tissues of CIN and CSCC patients, whose vaginal microecology was also altered. Both may play an important role in the progression of cervical lesions.

How Far Should We Investigate the Male Causes in Unexplained Infertility?

According to a recent report by the Guideline Development Group of the ESHRE, the definition of Unexplained Infertility (UI) is ‘infertility in couples with apparently normal ovarian function, fallopian tubes, uterus, cervix and pelvis, age [Formula: see text]40 years and with adequate coital frequency; and apparently normal testicular function, genito-urinary anatomy and a normal ejaculate.’ This is anchored with the ICMART definition of infertility, where couples should have at least 12 months of regular, unprotected sexual intercourse before investigations are initiated. Since Unexplained Infertility is a diagnosis by exclusion, apart from evaluating female causes, it seems important that in evaluating male contributions to the UI, all necessary tests need to be carried out. Unfortunately, there are no robust standardized methods for establishing etiologic diagnosis in the male partner and that in some patients, underlying sperm dysfunction may not be detected during a routine assessment of the ejaculate, which may appear to be normal using the 2021 WHO standards. It is possible that some adverse reproductive outcomes that have been identified during medical and reproductive history examination may require the use of further tests procedures specified in the latest WHO manual to assess sperm function. However, these tests have a research status only or are not regarded for routine use in clinical practice until sound evidence is developed. These tests include testicular imaging, anti-sperm antibodies, sperm DNA fragmentation, sperm chromatin condensation, sperm aneuploidy screening, serum hormone testing, semen HPV testing, and semen microbiology testing all of which were found not routinely indicated based on current evidence when semen was found to be normal using the WHO 2021 semen standards. In summary, in evaluating male causes in unexplained infertility, there are no robust data to support the recommendation that additional tests about the male reproductive function may routinely be carried out when semen analysis based on the 2021 WHO standard is normal. However, additional tests may be carried out when adverse reproductive outcomes and findings in the medical and reproductive history may require the use of these further tests especially when there are no findings on the female side.

MR Diffusion Kurtosis Imaging (DKI) of the Normal Human Uterus in Vivo During the Menstrual Cycle.

The uterus undergoes dynamic changes throughout the menstrual cycle. Diffusion kurtosis imaging (DKI) is based on the non-Gaussian distribution of water molecules and can perhaps represent the changes of uterine microstructure. To investigate the temporal changes in DKI-parameters of the normal uterine corpus and cervix during the menstrual cycle. Prospective. 21 healthy female volunteers (26.64 ± 4.72 years) with regular menstrual cycles (28 ± 7 days). Readout segmentation of long variable echo-trains (RESOLVE)-based DKI and fast spin-echo T2-weighted sequences at 3.0T. Each volunteer was scanned during the menstrual phase, ovulatory phase, and luteal phase. Regions of interest (ROI) were manually delineated in the endometrium, junctional zone, and myometrium of the uterine body, and in the mucosal layer, fibrous stroma layer, and loose stroma layer of the cervix. The mean Kapp (diffusion kurtosis coefficient), Dapp (diffusion coefficient), and ADC (apparent diffusion coefficient) values were measured in the ROI. ANOVA with Bonferroni or Tamhane correction. Intraclass correlation coefficient (ICC) for assessing agreement. P < 0.05 was considered statistically significant. During the menstrual cycle, the highest Kapp (0.848 ± 0.184) and lowest Dapp (1.263 ± 0.283 *10-3 mm2/sec) values were found in the endometrium during the menstrual phase. The Dapp values for the myometrium were significantly higher than those of the endometrium and the junctional zone in every phase. Meanwhile, the Dapp values for the three zonal structures of the cervix during ovulation were significantly higher than those during the luteal phase. However, there was no significant difference in the ADC values of the loose stroma between ovulation and the luteal phase (P = 0.568). The reproducibility of DKI parameters was good (ICC, 0.857-0.944). DKI can show dynamic changes of the normal uterus during the menstrual cycle. 2 TECHNICAL EFFICACY: Stage 2.

Role of Colposcopy in Diagnosing and Differentiating Types of Cervical Intraepithelial Neoplasia

Background: Preventing cervical cancer hinges on accurately detecting and treating high-grade cervical intraepithelial neoplasia (CIN2+) in individuals referred due to abnormal cervical cytology. While colposcopy has been a cornerstone in this process, concerns have arisen regarding its accuracy in CIN detection and differentiation. This study aims to assess the role of colposcopy in diagnosing and distinguishing various grades of CIN in clinically suspected patients. Methods: A descriptive cross-sectional study was conducted in the Department of Obstetrics &amp; Gynaecology at Rajshahi Medical College Hospital, Rajshahi, over 12 months from February 2019 to February 2020. The study involved 110 women aged 20-70 years who presented with clinical symptoms indicative of precancerous cervical lesions. Initial evaluations included visual inspection with 5% acetic acid (VIA) and Reid colposcopy. Biopsy samples were collected from the identified lesions using the colposcope, and these samples underwent histopathological examination. The study compared the diagnostic accuracy of colposcopy, including sensitivity, specificity, positive predictive value, and negative predictive value. Results: Colposcopic evaluations revealed that 80% of the participants exhibited positive lesions (CIN), while 20% displayed negative lesions (normal cervix or cervicitis). Among the colposcopically positive lesions, 50% were graded as CIN-1, 29.5% as CIN-2, and 20.5% as CIN-3. Subsequent histopathological examination of the colposcopy-directed biopsies confirmed CIN or dysplasia in 44.5% of cases. Within this group, 36.7% had CIN-1, 34.7% had CIN-2, 12.2% had CIN-3, 10.2% had invasive squamous cell carcinoma, and 6.2% had carcinoma in situ. Colposcopy demonstrated a sensitivity of 83.7% in diagnosing CIN, with a specificity of 23%, resulting in an overall diagnostic accuracy of 55%. However, when differentiating high-grade CIN from low-grade lesions, colposcopy exhibited a high sensitivity of 92.3% and moderate specificity of 67.7%, with an overall diagnostic accuracy of 75%. Conclusion: This study concludes that colposcopy is highly sensitive in diagnosing CIN but has limited specificity in excluding patients with normal cervix or cervicitis. Nevertheless, it exhibits optimal sensitivity in differentiating CIN2+ and carcinoma in situ from low-grade lesions and maintains moderate specificity in excluding low-grade CIN. Colposcopy remains a valuable tool in preventing and managing cervical cancer, particularly in identifying high-risk lesions.

Cervicovaginal microbiota disorder combined with the change of cytosine phosphate guanine motif- tolllike receptor 9 axis was associated with cervical cancerization.

Convincing studies demonstrated that cervicovaginal microbiota disorder and toll-like receptor 9 (TLR9) high expression were related to cervical carcinogenesis. However, the effects of cervicovaginal microbiota integration TLR9 in cervical cancerization are unclear. Based on the biological basis that unmethylated cytosine-phosphate-guanine (CpG) motifs of bacteria could activate TLR9, we explored the effects of cervicovaginal microbiota disorder and CpG motif-TLR9 axis change in cervical carcinogenesis. A total of 341 participants, including 124 normal cervical (NC), 90 low-grade cervical intraepithelial neoplasia (CIN1), 78 high-grade cervical intraepithelial neoplasia (CIN2/3) and 49 squamous cervical cancer (SCC), diagnosed by pathology were enrolled in the study. Here, metagenomic shotgun sequencing was used to reveal cervicovaginal microbiota characteristics, and TLR9 protein was detected by western blotting. Our results showed that the diversity of cervicovaginal microbiota gradually increased along with the poor development of cervical lesions, showing the abundance of Lactobacillus crispatus and Lactobacillus iners decreased, while the abundance of pathogenic bacteria gradually increased. The level of TLR9 expression was gradually increased with cervicovaginal microbiota diversity increasing, the abundance of Lactobacillus decreasing, and we found a positive correlation dependency relationship (r = 0.384, P = 0.002) between TLR9 and GTCGTT motif content. Stratified analysis based on HPV16 infection, we found that the characteristics of cervicovaginal microbiota and increased TLR9 expression were also closely related to HPV16 infection. Cervicovaginal microbiota dysbiosis might lead to the CpG motif increased, which was closely associated with TLR9 high expression, and ultimately might promote the progression of cervical lesions.

Diverse ultrasound image features of unilateral genital tract obstruction with ipsilateral renal anomaly syndrome on genitourinary system segmental sequential ultrasound screening and the accuracy of ultrasonic diagnosis.

Unilateral genital tract obstruction with ipsilateral renal anomaly (UGTOIRA) syndrome is a rare congenital urogenital anomaly, characterized by different combinations of uterine abnormalities, unilateral cervical-vaginal obstruction, and ipsilateral renal abnormalities. Timely and correct diagnosis is critical. In this study, we analyzed the diverse ultrasound image features of UGTOIRA syndrome on genitourinary system segmental sequential ultrasound screening (SSUS) and the accuracy of ultrasonic diagnosis. The data from 59 patients with UGTOIRA syndrome over the last decade were analyzed retrospectively, which included ultrasound presentations and diagnoses of abnormalities of the uterus, cervix, and vagina, as well as the conditions associated with the bilateral fallopian tubes, ovaries, pelvis, kidneys and ureters, bladder, and urethra. All 59 patients (100%) were found to have ipsilateral renal agenesis, and the diagnostic accuracy of ultrasound was 100%. The uterus was correctively diagnosed as complete bicorporal uterus in 42 cases (71.2%), bicorporal septate uterus in 7 cases (11.9%), complete uterus septate in 8 cases (13.6%), partial uterus septate in 1 case (1.7%), and unilateral isthmus atresia in 1 case (1.7%). The cervix was correctively diagnosed as septate cervix in 28 cases (47.5%), double cervix in 17 cases (28.8%), unilateral small cervix in 5 cases (8.5%), unilateral obliterated cervical orifice in 5 cases (8.5%), partial development of the unilateral cervix with a blind end in 3 cases (5.1%), and unilateral completely undeveloped cervix appearing as a normal cervix in 1 case (1.7%). The vagina was correctively diagnosed as oblique vaginal septum (OVS) in 45 cases (76.3%), and normal vagina in 13 cases (22.0%) via preoperative ultrasound. In 1 case (1/59, 1.7%) of high OVS with ipsilateral obliterated external cervical orifice, the OVS was missed by preoperative ultrasound. The ultrasound diagnostic accuracy rates of uterine malformation, cervical malformation, vaginal malformation, hematoma location and volume, uterine communication, and cervical communication were respectively 100%, 100%, 98.3%, 100%, 100%, and 100%. The ultrasonic detection rate for a hole on the OVS was 0%, and the missed diagnosis rate was 100%. SSUS can be used to accurately evaluate uterine, cervical, and vaginal malformation, obstruction site, location and volume of keratocele, cervical or uterine communication, and ovarian endometriosis cyst in patients with UGTOIRA syndrome and ipsilateral urinary system malformation. However, ultrasound is unable to identify a hole on the OVS.

Assessing the capacity of methylated DNA markers of cervical squamous cell carcinoma to discriminate oropharyngeal squamous cell carcinoma in human papillomavirus mediated disease

ObjectiveEarly identification of human papillomavirus associated oropharyngeal squamous cell carcinoma (HPV(+)OPSCC) is challenging and novel biomarkers are needed. We hypothesized that a panel of methylated DNA markers (MDMs) found in HPV(+) cervical squamous cell carcinoma (CSCC) will have similar discrimination in HPV(+)OPSCC tissues. Materials and methodsFormalin-fixed, paraffin-embedded tissues were obtained from patients with primary HPV(+)OPSCC or HPV(+)CSCC; control tissues included normal oropharynx palatine tonsil (NOP) and cervix (NCS). Using a methylation-specific polymerase chain reaction, 21 previously validated cervical MDMs were evaluated on tissue-extracted DNA. Discrimination between case and control cervical and oropharynx tissue was assessed using area under the curve (AUC). Results34 HPV(+)OPSCC, 36 HPV(+)CSCC, 26 NOP, and 24 NCS patients met inclusion criteria. Within HPV(+)CSCC, 18/21 (86%) of MDMs achieved an AUC ≥ 0.9 and all MDMs exhibited better than chance classifications relative to control cervical tissue (all p < 0.001). In contrast, within HPV(+)OPSCC only 5/21 (24%) MDMs achieved an AUC ≥ 0.90 but 19/21 (90%) exhibited better than chance classifications relative to control tonsil tissue (all p < 0.001). Overall, 13/21 MDMs had statistically significant lower AUCs in the oropharyngeal cohort compared to the cervical cohort, and only 1 MDM exhibited a statistically significant increase in AUC. ConclusionsPreviously validated MDMs exhibited robust performance in independent HPV(+)CSCC patients. However, most of these MDMs exhibited higher discrimination for HPV(+)CSCC than for HPV(+)OPSCC. This suggests that each SCC subtype requires a unique set of MDMs for optimal discrimination. Future studies are necessary to establish an MDM panel for HPV(+)OPSCC.

Differential expression of mRNA 3′-end isoforms in cervical and ovarian cancers

Early diagnosis and therapeutic targeting are continuing challenges for gynecological cancers. Here, we focus on cancer transcriptomes and describe the differential expression of 3′UTR isoforms in patients using an algorithm to detect differential poly(A) site usage. We find primarily 3′UTR shortening cases in cervical cancers compared with the normal cervix. We show differential expression of alternate 3′-end isoforms of FOXP1, VPS4B, and OGT in HPV16-positive patients who develop high-grade cervical lesions compared with the infected but non-progressing group. In contrast, in ovarian cancers, 3′UTR lengthening is more evident compared with normal ovary tissue. Nevertheless, highly malignant ovarian tumors have unique 3′UTR shortening events (e.g., CHRAC1, SLC16A1, and TOP2A), some of which correlate with upregulated protein levels in tumors. Overall, our study shows isoform level deregulation in gynecological cancers and highlights the complexity of the transcriptome. This transcript diversity could help identify novel cancer genes and provide new possibilities for diagnosis and therapy.

Potential diagnostic value of quantitative superb microvascular imaging in premalignant and malignant cervical lesions.

The purpose of this study was to assess the diagnostic efficacy of the vascular index (VI) on superb microvascular imaging (SMI) in distinguishing normal uterine cervical epithelium, high-grade cervical intraepithelial neoplasia (CIN), and cervical cancer. The retrospective study included women with pathology-confirmed CIN or cervical cancer, who underwent transvaginal ultrasound and SMI between April 2021 and October 2022. The SIM manifestations of normal cervix and cervical lesions were reviewed. SIM were measured and converted into vascular index (VI) which compared between cervical lesions and control groups. We have retrospectively compared ultrasound features of cervical lesions and characteristics of patients. Measurement reliability was evaluated by intra class correlation coefficient (ICC). A total of 235 consecutive females were enrolled, comprising 38 with high-grade CIN, 96 with cervical cancer, and 101 with a normal uterine cervix. The microvascular architecture exhibited significant variations between premalignant and malignant cervical lesions. Branch-like patterns were predominantly observed in high-grade CIN, while crab claw-like and fireball-like patterns were more commonly associated with cervical cancer. The median VI of cervical cancer (34.7 ± 10.3) was significantly higher than that of high-grade CIN (17.6 ± 4.2) (P < 0.001). Moreover, the VI values of cervical cancer differed significantly among different FIGO stages and pathological types (P < 0.001 and P = 0.003, respectively). The VI demonstrated superior diagnostic performance for cervical lesions compared to vascular patterns (AUC = 0.974 and 0.969, respectively). Using a cut-off value of 25.5, the VI yielded a sensitivity of 82.3% and a specificity of 99.3% for cervical lesion detection. The SMI parameter (VI) exhibited a significantly higher value in cervical cancer compared to high-grade CIN, with a high level of agreement among observers. These findings suggest that quantitative SMI holds promise as an imaging technique for the detection and characterization of cervical lesions.

TIME Is Ticking for Cervical Cancer.

Cervical cancer (CC) is a major health problem among reproductive-age females and comprises a leading cause of cancer-related deaths. Human papillomavirus (HPV) is the major risk factor associated with CC incidence. However, lifestyle is also a critical factor in CC pathogenesis. Despite HPV vaccination introduction, the incidence of CC is increasing worldwide. Therefore, it becomes critical to understand the CC tumor immune microenvironment (TIME) to develop immune cell-based vaccination and immunotherapeutic approaches. The current article discusses the immune environment in the normal cervix of adult females and its role in HPV infection. The subsequent sections discuss the alteration of different immune cells comprising CC TIME and their targeting as future therapeutic approaches.

Cervical carcinoma induces NLRP3 inflammasome activation and IL-1ß release in human peripheral blood monocytes affecting patients' overall survival.

Our group previously demonstrated that genetic variants in inflammasome genes contribute to protection against the establishment of human papilloma virus (HPV)-associated cervical carcinoma (CC). The objective of this study was to better understand the contribution of inflammasome and its cytokines in the CC microenvironment. The inflammasome activation was analyzed in CC tumoral cell lines and healthy donors (HD)' monocytes in co-culture. In vitro results were then compared to CC patients' public databases. CC cells did not produce IL-1ß or IL-18 themselves, however, when in co-culture with HD monocytes, induced IL-1ß release in those leucocytes. Inflammasome activation appears to be partially dependent on the NLRP3 receptor. Public data analysis revealed that IL1B expression is increased in the CC compared to normal uterine cervix, and that patients with high IL1B expression had a shorter overall survival. CC microenvironment can activate the inflammasome and IL-1ß release in surrounding monocytes, which could be detrimental for CC prognosis.

High-resolution reduced field-of-view diffusion-weighted magnetic resonance imaging in the diagnosis of cervical cancer.

Magnetic resonance imaging (MRI) has now become the best modality for the preoperative staging of cervical cancer. This study was to compare the value of high-resolution reduced field-of-view diffusion-weighted MR imaging (r-FOV DWI) with conventional field-of-view (c-FOV DWI) in the diagnosis of cervical cancer. Forty-five patients (25 patients with cervical cancer and 20 patients with normal cervix) received magnetic resonance (MR) scans (3.0T), including both r-FOV DWI and c-FOV DWI sequences. The image quality (IQ) of both sequences was subjectively assessed by two attending radiologists using a double-blind method and quantitatively by the signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR). Moreover, apparent diffusion coefficient (ADC) values for cervical cancer were blindly measured by one technician on the ADC map. The subjective scores of r-FOV DWI images were higher than those of c-FOV DWI (P<0.0001), and the interrater reliability was in good agreement [Cohen's kappa coefficient (κ) =0.547-0.914]. There was a significant difference in CNR between the two DWI image groups (r-FOV DWI 12.73±5.56 vs. c-FOV DWI 11.21±5.92, P=0.019). The difference in mean ADC values between the two DWI sequences was statistically significant [r-FOV DWI (0.690±0.195)×10-3 mm2/s vs. c-FOV DWI (0.794±0.167)×10-3 mm2/s, P<0.001]. The ADC value of cervical cancer lesions [(0.690±0.195)×10-3 mm2/s] was significantly lower than that of normal cervix ADC value [(1.506±0.188)×10-3 mm2/s]. r-FOV DWI can effectively improve the spatial resolution of the image while reducing distortion and artifacts. Furthermore, it can help to diagnose cervical cancer more accurately for the more realistic ADC values.