Proteomic detection of COX-2 pathway-related factors in patients with adenomyosis.

Background: Investigating the relationship between cyclooxygenase-2 (COX-2) pathway-related factors and clinical features in patients with adenomyosis by proteomics could provide potential therapeutic targets. Methods: This study recruited 40 patients undergoing surgical hysterectomy and pathological diagnosis of adenomyosis, collected ectopic endometrial specimens, and recorded clinical data. The expression levels of COX-2 in ectopic uterus lesions were detected using the immunohistochemical (IHC) SP method. The 40 samples were then divided into a COX-2 low or high expression group. Five samples with the most typical expression levels were selected from each of the two groups and the differential proteins between the two groups were identified using label-free quantitative proteomics. WW domain-binding protein 2 (WBP2), interferon induced transmembrane protein 3 (IFITM3), and secreted frizzled-related protein 4 (SFRP4) were selected for further verification, and their relationships with COX-2 and clinical characteristics were analyzed. Results: There were statistically significant differences in the expression of WBP2, IFITM3, and SFRP4 between the COX-2 low and high expression groups (P < 0.01). The expressions of COX-2, IFITM3, and SFRP4 were significantly correlated with dysmenorrhea between the two groups (P < 0.05), but not with uterine size or menstrual volume (P > 0.05). However, there was no significant correlation between the expression of WBP2 and dysmenorrhea, uterine size, and menstruation volume in both the high expression and low expression groups (P > 0.05). Conclusions: COX-2, IFITM3, SFRP4, and WBP2 may be involved in the pathogenesis of adenomyosis. COX-2, IFITM3, and SFRP4 may serve as potential molecular biomarkers or therapeutic targets in dysmenorrhea in patients with early adenomyosis.

Esomeprazole Alleviates Cisplatin Resistance by Inhibiting the AKT/mTOR Pathway in Ovarian Cancer Cells.

Purpose: Ovarian cancer is the most lethal malignancy in gynecology. Due to limited treatment strategies and platinum resistance, newer drugs and therapeutic options are needed. Esomeprazole (ESO) has been reported to have multiple anticancer activities in preclinical and clinical research. Therefore, this study aimed to explore the anticancer effects of esomeprazole on ovarian cancer and its underlying molecular mechanisms. Methods: CCK-8 and 5-ethynyl-2'-deoxyuridine (EdU) assays were used to detect cell viability and proliferation. The Transwell assay was used to evaluate cell migration and invasion capacity. Flow cytometry was used to detect cell apoptosis. Western blotting and immunofluorescence were used to detect protein expression. Results: ESO effectively inhibited the cell viability, proliferation, invasion, migration, and induced apoptosis of ovarian cancer cells in a concentration-dependent manner. Treatment with ESO decreased the expression of c-MYC, SKP2, E2F1, N-cadherin, vimentin, and matrix metalloproteinase 2 (MMP2), while it increased E-cadherin, caspase3, p53, BAX, and cleaved poly (ADP-ribose) polymerase (PARP) expression, and downregulated the PI3K/AKT/mTOR signaling pathway. Furthermore, ESO combined with cisplatin showed synergistic effects in inhibiting proliferation, invasion, and migration of cisplatin-resistant ovarian cancer cells. The mechanism may be related to the increased inhibition of c-MYC, epithelial-mesenchymal transition (EMT), and the AKT/mTOR signaling pathway and enhanced the upregulation of the pro-apoptotic protein BAX and cleaved PARP levels. Moreover, ESO combined with cisplatin synergistically upregulated the expression of the DNA damage marker γH2A.X. Conclusion: ESO exerts multiple anticancer activities and has a synergistic effect in combination with cisplatin on cisplatin-resistant ovarian cancer cells. This study provides a promising strategy to improve chemosensitivity and overcome resistance to cisplatin in ovarian cancer.

Fine-grained Lesion Classification Framework for Early Auxiliary Diagnosis.

The deep neural networks are envisaged for the early disease diagnosis from medical images. However, in the early stage of the disease, the medical images of patients and healthy people have only subtle visual differences. Distinguishing the medical images for early diagnosis belongs to the Fine-Grained Visual Classification (FGVC) task. Many recent works are based on a standard FGVC learning paradigm: locate the discriminative regions first and then classify by fusing the information of these regions. However, it is still not enough for medical images. Because the shape and size of the lesions are variable, and the relationship between lesions and the background is complex. In order to solve these problems, we propose a fine-grained lesion classification framework for early auxiliary diagnosis. We first locate and extract multiple lesions with different sizes and shapes from the original image and then fuse the feature of lesion and background based on attention mechanism. As shown by experiment results in two real-world clinical data sets, our model can locate accurately and perform better.

Vaginal yolk sac tumors in infants and children: A case report and literature review.

A vaginal yolk sac tumor (YST) is a rare malignant germ cell tumor for infants and children, and it has been >50 years since the first case was reported. The treatment strategy has changed markedly in the past 50 years, from radical surgical treatment to conservative surgery combined with chemotherapy, and then to combined chemotherapy alone. The present study reports the case of a primary vaginal YST in a 13-month-old girl that was successfully treated by tumor resection combined with chemotherapy. The clinical symptoms, imaging features and treatment characteristics are described in detail, as well as the postoperative treatment. There was no local recurrence or metastasis for the 2 years of follow-up to date. A literature review was also conducted to investigate the clinicopathological features, treatment and prognosis of this tumor. Overall, surgery combined with bleomycin, etoposide and carboplatin combination chemotherapy can be an effective option for vaginal YST.

Celecoxib reduces inflammation and angiogenesis in mice with adenomyosis.

OBJECTIVE: This study aimed to explore the effect of COX-2 selective inhibitor (celecoxib) on adenomyosis and its mechanism. METHODS: By establishing a mouse model of adenomyosis and using celecoxib to treat adenomyosis, newly born female mice were randomly divided into a control group, adenomyosis model group, and celecoxib group. Hematoxylin-eosin (H&E) staining was used to observe the depth of endometrial infiltration of mouse adenomyosis. RT-PCR (reverse transcription PCR) and western blot were used to detect the expression of Cyclooxygenase-2 (COX-2), Vascular growth factor (VEGF), Nerve growth factor (NGF), and Corticotropin-releasing hormone (CRH) mRNA and protein in mice before and after celecoxib treatment. RESULTS: After treatment with celecoxib, the depth of endometrial infiltration of mouse adenomyosis was reduced. COX-2 and VEGF decreased significantly after celecoxib inhibited expression of COX-2 (P<0.001), but there was no significant difference in the expression of NGF or CRH (P>0.05). CONCLUSION: This study indicated that COX-2 may be an important factor related to the pathogenesis of adenomyosis, and it may become an important molecular target for the treatment of adenomyosis.

Identification and characterization of a murine model of BCR­ABL1+ acute B­lymphoblastic leukemia with central nervous system metastasis.

Breakpoint cluster region (BCR)­Abelson murine leukemia (ABL)1+ acute B­lymphoblastic leukemia (B­ALL) is a disease associated with a dismal prognosis and a high incidence of central nervous system (CNS) metastasis. However, BCR­ABL1+ B­ALL with CNS infiltration has not been previously characterized, at least to the best of our knowledge. In the present study, a murine model of BCR­ABL1+ B­ALL with CNS metastasis was established using retroviral transduction. The vast majority of BCR­ABL1+ leukemic cells were found to be immature B cells with a variable proportion of pro­B and pre­B populations. The present results indicated that the BCR­ABL1+ B­leukemic cells expressed high levels integrin subunit alpha 6 (Itga6) and L­selectin adhesion molecules, and have an intrinsic ability to disseminate and accumulate in CNS tissues, predominantly in meninges. On the whole, these results provide an approach for addressing the mechanisms of BCR­ABL1+ B­ALL with CNS metastasis and may guide the development of novel therapeutic strategies.

Computer-Aided Diagnosis of Label-Free 3-D Optical Coherence Microscopy Images of Human Cervical Tissue.

OBJECTIVE: Ultrahigh-resolution optical coherence microscopy (OCM) has recently demonstrated its potential for accurate diagnosis of human cervical diseases. One major challenge for clinical adoption, however, is the steep learning curve clinicians need to overcome to interpret OCM images. Developing an intelligent technique for computer-aided diagnosis (CADx) to accurately interpret OCM images will facilitate clinical adoption of the technology and improve patient care. METHODS: 497 high-resolution three-dimensional (3-D) OCM volumes (600 cross-sectional images each) were collected from 159 ex vivo specimens of 92 female patients. OCM image features were extracted using a convolutional neural network (CNN) model, concatenated with patient information [e.g., age and human papillomavirus (HPV) results], and classified using a support vector machine classifier. Ten-fold cross-validations were utilized to test the performance of the CADx method in a five-class classification task and a binary classification task. RESULTS: An 88.3 ± 4.9% classification accuracy was achieved for five fine-grained classes of cervical tissue, namely normal, ectropion, low-grade and high-grade squamous intraepithelial lesions (LSIL and HSIL), and cancer. In the binary classification task [low-risk (normal, ectropion, and LSIL) versus high-risk (HSIL and cancer)], the CADx method achieved an area-under-the-curve value of 0.959 with an 86.7 ± 11.4% sensitivity and 93.5 ± 3.8% specificity. CONCLUSION: The proposed deep-learning-based CADx method outperformed four human experts. It was also able to identify morphological characteristics in OCM images that were consistent with histopathological interpretations. SIGNIFICANCE: Label-free OCM imaging, combined with deep-learning-based CADx methods, holds a great promise to be used in clinical settings for the effective screening and diagnosis of cervical diseases.

Ultrahigh-resolution optical coherence microscopy accurately classifies precancerous and cancerous human cervix free of labeling.

Cervical cancer remains the fourth most common cause of cancer worldwide and the third leading cause of cancer deaths for women in developing countries. Traditional screening tools, such as human papillomavirus and Pap tests, cannot provide results in real-time and cannot localize suspicious regions. Colposcopy-directed biopsies are invasive in nature and only a few sites of the cervix may be chosen for investigation. A non-invasive, label-free and real-time imaging method with a resolution approaching that of histopathology is desirable for early detection of the disease. Methods: Ultrahigh-resolution optical coherence microscopy (OCM) is an emerging imaging technique used to obtain 3-dimensional (3-D) "optical biopsies" of biological samples with cellular resolution. In this study, 497 3-D OCM datasets from 159 specimens were collected from 92 patients. Results: Distinctive patterns for normal cervix, squamocolumnar junction, ectropion, low-grade and high-grade squamous intraepithelial lesions (LSIL and HSIL) and invasive cervical lesions were clearly observed from OCM images, which matched well with corresponding histological slides. OCM images demonstrated a sensitivity of 80% (95% confidence interval, CI, 72%-86%) and a specificity of 89% (95% CI, 84%-93%) for detecting high-risk lesions (HSIL and invasive lesions) when blindly tested by three investigators. A substantial inter-observer agreement was observed (κ=0.627), which showed high diagnostic consistency among three investigators. Conclusion: These results laid the foundation for future non-invasive optical evaluation of cervical tissue in vivo, which could lead to a less invasive and more effective screening and "see-and-treat" strategy for the management of cervical cancer.

[Targeted interruption of COX-2 gene by siRNA inhibits the expression of VEGF, MMP-9, the activity of COX-2 and stimulates the apoptosis in eutopic, ectopic endometrial stromal cells of women with endometriosis].

OBJECTIVE: To investigate the effect of targeted interruption of cyclooxygenase-2 (COX-2) gene by small interference RNA (siRNA) on the expression of COX-2, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in eutopic and ectopic endometrial stromal cells (ESC) with endometriosis, and the effect on the content of 6-keto-prostaglandin-F1α (6-keto-PGF1α, metabolites of COX) and the apoptosis of eutopic and ectopic ESC with endometriosis. METHODS: Ectopic and eutopic ESC from 30 women with endometriosis were isolated and cultured respectively. Then, ESC were classified into three groups: interference group, negative control group and blank control group. ESC in interference group were injected into siRNA transfection complex while ESC in negative control group were injected into negative control transfection complex. ESC from 10 participants without endometriosis were the normal control group. The mRNA and protein expression of COX-2, VEGF, MMP-9 in pre-transfected and post-transfected eutopic and ectopic ESC were detected through real time reverse transcription PCR and western blot. The content of 6-keto-PGF1α was determined by ELISA, the apoptotic cells were detected by flow cytometry. RESULTS: After interruption of COX-2 gene, there were no significant difference in the mRNA and protein expression of COX-2, VEGF and MMP-9 between the negative control group and blank control group (P > 0.05); the mRNA and protein expression of the three genes in interference group were significantly lower than those in negative control group and blank control group (P < 0.05); the mRNA expression of the three genes in interference group of eutopic ESC were 0.87 ± 0.06, 1.76 ± 0.59, 1.04 ± 0.32, in interference group of ectopic ESC were 0.75 ± 0.12, 1.62 ± 0.47, 0.88 ± 0.25, the protein expression of the three genes in interference group of eutopic ESC were 0.457 ± 0.019, 0.500 ± 0.012, 0.361 ± 0.008, in interference group of ectopic ESC were 0.323 ± 0.018, 0.474 ± 0.016, 0.339 ± 0.009; the mRNA and protein expression of the three genes in ectopic ESC had a more reduction than those in eutopic ESC (P < 0.05). The results from ELISA revealed that the content of 6-keto-PGF1α in the normal control group [(17.7 ± 1.9) pg/ml] were significantly lower than those in the blank control group (P < 0.05), the content of 6-keto-PGF1α in ectopic ESC were significantly higher than that in eutopic ESC (P < 0.05), the content of 6-keto-PGF1α in the blank control group of eutopic and ectopic ESC were (32.4 ± 2.6) pg/ml, (38.2 ± 3.7) pg/ml; there was no significant difference in the content of 6-keto-PGF1α between the negative control group and blank control group (P > 0.05); compared with those of negative control group and blank control group, the content of 6-keto-PGF1α in interference group decreased significantly (P < 0.05), the content of 6-keto-PGF1α in interference group of eutopic and ectopic ESC were (17.1 ± 2.4) pg/ml, (20.9 ± 2.7) pg/ml; the content of 6-keto-PGF1α in eutopic ESC had a slightly more reduction than that in ectopic ESC (P > 0.05). The results from flow cytometry displayed that, there was no significant difference in apoptotic cells between the negative control group and blank control group (P > 0.05); compared with those of negative control group and blank control group, more apoptotic cells were detected in interference group and the difference was significant (P < 0.01); the apoptotic cells in ectopic ESC were significantly more than that in eutopic ESC (P < 0.05); the apoptosis rate in interference group of eutopic and ectopic ESC were (33.76 ± 0.06)%, (47.18 ± 0.12)%. CONCLUSIONS: Our results suggested the targeted interruption of COX-2 gene by siRNA effectively inhibited the mRNA and protein expression of COX-2, VEGF and MMP-9 in both eutopic ESC and ectopic ESC with endometriosis, greatly increased the apoptotic rate of cells and obviously reduced the content of 6-keto-PGF1α by inhibiting the activity of COX-2. And the changes in ectopic endometrium were more evident than those in eutopic endometrium.

Diagnostic value of serum CA125, CA19-9 and CA15-3 in endometriosis: A meta-analysis.

AIM: To evaluate the diagnostic value of serum cancer antigen (CA)125, CA19-9 and CA15-3 concentrations in endometriosis. METHODS: Case-control studies evaluating CA125, CA19-9 and CA15-3 and endometriosis, published between January 2000 and November 2014 were retrieved from PubMed(®) and Google Scholar. Standardized mean differences (SMD) and 95% confidence intervals (CI) were calculated. Subgroup analyses were carried out by ethnicity and disease stage (early, stage I/II; advanced, stage III/IV). RESULTS: The analysis included 12 case-control studies (963 cases, 855 controls). CA125 was associated with endometriosis in the overall population (SMD 0.82, 95% CI 0.72, 0.92), Caucasian subgroup (SMD 1.08, 95% CI 0.96, 1.19), and early (SMD 1.20, 95% CI 0.93, 1.48) or advanced disease (SMD 1.29, 95% CI 1.04, 1.55). CA19-9 was associated with endometriosis in the overall population (SMD 0.48, 95% CI 0.24, 0.72), Caucasian subgroup (SMD 0.31, 95% CI 0.07, 0.55), Asian subgroup (SMD 9.65, 95% CI 7.88, 11.42) and advanced disease (SMD 0.60, 95% CI 0.34, 0.87). CA15-3 was significantly associated with advanced disease (SMD 0.47, 95% CI 0.09, 0.84). CONCLUSIONS: Serum CA125 and CA19-9 may represent useful biomarkers for the noninvasive diagnosis of endometriosis.

[Correlation between P53 codon 72 polymorphism and tumorigenesis of cervical cancer].

BACKGROUND & OBJECTIVE: The p53 codon 72 polymorphism affects human papillomavirus (HPV) E6-mediated degradation of p53. This study was to investigate distribution of p53 polymorphism in Guangdong women, and relationship between p53 polymorphism and tumorigenesis of cervical cancer. METHODS: Cervical smears of 46 patients with cervical cancer (case group), and 84 patients with benign gynecologic tumor (control group) treated in our hospital from Sept. 2002 to May 2003 were collected. DNA, extracted from cervical smears,were examined by polymerase chain reaction (PCR) for detection of HPV DNA and p53 codon 72 polymorphism. RESULTS: Positive rate of HPV DNA in case group was 47.8%, in control group was 20.2%. Proportions of genotypes Arg/Arg, Pro/Pro, and Arg/Pro in case group were 56.5%, 21.7%,and 21.7%, respectively; in control group were 71.4%,20.2%, and 8.3%,respectively. There were no significant differences in proportions of Arg/Arg (OR,0.520; 95% CI,0.245-1.102), and Arg/Pro (OR, 1.095; 95% CI,0.454-2.639) between 2 groups; proportion of Pro/Pro in case group was significantly higher than control group (OR, 3.056; 95% CI,1.076-8.678), but no significant difference was found in women with HPV infection. CONCLUSION: Arg/Arg genotype is not a high-risk factor for cervical cancer in Chinese population, and individuals with Pro/Pro genotype are likely to develop cervical cancer.