PAX2 is regulated by estrogen/progesterone through promoter methylation in endometrioid adenocarcinoma and has an important role in carcinogenesis via the AKT/mTOR signaling pathway.

Endometrioid adenocarcinoma (EEC) is one of the most common cancers of the female reproductive system. In recent years, much emphasis has been placed on early diagnosis and treatment. PAX2 (Paired box 2) inactivation is reportedly an important biomarker for endometrioid intraepithelial neoplasia (EIN) and EEC. However, the role of PAX2 in EEC carcinogenesis remains unclear. PAX2 expression and associated clinical characteristics were analyzed via The Cancer Genome Atlas, Gene Expression Omnibus, and Cancer Cell Line Encyclopedia databases and clinical paired EIN/EEC tissue samples. Bioinformatic analysis was conducted to identify the putative molecular function and mechanism of PAX2. Cell proliferation, colony formation, cell migration, and invasion assays in vitro, and mouse xenograft models were utilized to study the biological functions of PAX2 in vivo. Pyrosequencing and the demethylating drug 5-Aza-dc were used to verify promoter methylation in clinical tissues and cell lines, respectively. The mechanism underlying the regulatory effect of estrogen (E2) and progesterone (P4) on PAX2 expression was investigated by receptor block assay and double luciferase reporter assay. PAX2 expression was found to be significantly downregulated in EIN and EEC tissues, its overexpression inhibited EEC cell malignant behaviors in vivo and in vitro and inhibited the AKT/mTOR signaling pathway. PAX2 inactivation in EEC was related to promoter methylation, and its expression was regulated by E2 and P4 through their receptors via promoter methylation. Our findings elucidated the expression and function of PAX2 in EEC and have provided hitherto undocumented evidence of the underlying molecular mechanisms. PAX2 expression is suppressed by estrogen prompting its methylation through estrogen receptor. Furthermore, PAX2 regulates the AKT/mTOR signaling pathway to influence EEC progression. © 2024 The Pathological Society of Great Britain and Ireland.

TFE3 nuclear expression as a novel biomarker of ovarian sclerosing stromal tumors and associated with its histological morphology.

Sclerosing stromal tumors of the ovary are benign and tend to occur in youthful women with lobular structures at low frequencies. Three types of cells, including luteinized cells, short spindle myoid cells, and intermediate cells, are found in the lobules which abundant in the blood vessels. Currently, immunohistochemistry is used to detect normal follicles, sclerosing stromal tumors, granulosa cell tumors, and fibromas/thecomas. Our research results showed that transcription factor enhancer 3 (TFE3) was moderate to strong positive in the theca interna layer of normal follicles. TFE3 was expressed in seven out of eight sclerosing stromal tumors, mainly in luteinized cells. It did not express in 20 granulosa cell tumors. Of the nine fibromas/thecomas, TFE3 was weakly staining in 2 cases and negative in the remaining 7 cases. The expression of TFE3 was also weak in only one microcystic stromal tumor. 8 cases of sclerosing stromal tumors were analyzed by FISH using a TFE3 separation probe, and the results were negative. In short, as a nuclear transcription protein, TFE3 specifically expressed in sclerosing stromal tumors and could serve as a new marker for the diagnosis and differential diagnosis of sclerosing stromal tumors. Moreover, we speculate that TFE3 will promotes the formation of the vascular plexus after entry into the nucleus, which can further explain why sclerosing stromal tumors are different from other ovary sex-cord stromal tumors.

CUL4A-mediated ZEB1/microRNA-340-5p/HMGB1 axis promotes the development of osteoporosis.

Understanding the molecular mechanisms underlying osteoclast differentiation provides insights into bone loss and even osteoporosis. The specific mechanistic actions of cullin 4A (CUL4A) in osteoclast differentiation and resultant osteoporosis is poorly explored. We developed a mouse model of osteoporosis using bilateral ovariectomy (OVX) and examined CUL4A expression. It was noted that CUL4A expression was increased in the bone marrow of OVX mice. Overexpression of CUL4A promoted osteoclast differentiation, and knockdown of CUL4A alleviated osteoporosis symptoms of OVX mice. Bioinformatic analyses were applied to identify the downstream target genes of microRNA-340-5p (miR-340-5p), followed by interaction analysis. The bone marrow macrophages (BMMs) were isolated from femur of OVX mice, which were transfected with different plasmids to alter the expression of CUL4A, Zinc finer E-box binding homeobox 1 (ZEB1), miR-340-5p, and Toll-like receptor 4 (TLR4). ChIP assay was performed to detect enrichment of ZEB1 promoter by H3K4me3 antibody in BMMs. ZEB1 was overexpressed in the bone marrow of OVX mice. Overexpression of CUL4A mediated H3K4me3 methylation to increase ZEB1 expression, thus promoting osteoclast differentiation. Meanwhile, ZEB1 could inhibit miR-340-5p expression and upregulate HMGB1 to induce osteoclast differentiation. Overexpressed ZEB1 activated the TLR4 pathway by regulating the miR-340-5p/HMGB1 axis to induce osteoclast differentiation, thus promoting the development of osteoporosis. Overall, E3 ubiquitin ligase CUL4A can upregulate ZEB1 to repress miR-340-5p expression, leading to HMGB1 upregulation and the TLR4 pathway activation, which promotes osteoclast differentiation and the development of osteoporosis.

CIN grades possessing different HPV RNA location patterns and RNAscope is helpful tool for distinguishing squamous intraepithelial lesions in difficult cervical cases.

BACKGROUND AND OBJECTIVES: The precise grading and characterization of cervical intraepithelial neoplasia (CIN) has been the focus of pathologists for a long time. This study aimed to explore known strategies for the grading of CINs. METHODS: After routine H&E review, 85 lesions graded CIN 1, 2, or 3 were investigated primarily by HPV RNAscope to detect HR-HPV and LR-HPV, in combination with an HPV-DNA test and P16/Ki67 immunohistochemistry (IHC). Then, the 85 cases were divided into a control group (49 cases) and a test group (36 cases). The former consisted of cases with consistency between morphology, HPV DNA detection and P16/Ki67 IHC. We used them to evaluate HPV RNA distribution patterns in CINs of different grades. The latter were ambiguous cases in which pathologists could not confirm the diagnosis because of inconsistencies between morphology, HPV DNA detection and P16/Ki67 IHC. We reassessed them by comparison to the pattern in the control group. RESULTS: The expression patterns of HPV mRNA signals were different in different CIN lesions. LSIL/CIN1 lesions were mostly expressed in superficial epithelium with diffuse clustered nuclear or cytoplasmic staining; HSIL/CIN2 were characterised by nuclear/cytoplasmic punctate or diffuse cluster nuclear staining in the mid-surface layer, and scattered nuclear/cytoplasmic punctate staining in basal and parabasal cells; whereas HSIL/CIN3 showed full-thickness nucleus/cytoplasmic scattered staining with a punctate pattern. According to the staining pattern, we corrected the diagnosis of 22 cases (22/36, 61.1%). CONCLUSION: Because of its distinct location pattern, HPV RNAscope has obvious advantages over the HPV-DNA test, and combined with P16/Ki67 IHC, it can help pathologists correctly grade CIN. In addition, it can effectively discriminate true CIN from normal or CIN mimic lesions, such as immature squamous metaplasia, atrophy, and inflammatory/reactive changes. Therefore, HPV RNAscope is a valuable auxiliary diagnostic test to avoid the overtreatment and undertreatment of CIN lesions.

Fluorine-functionalized conjugated microporous polymer as adsorbents for solid-phase extraction of nine perfluorinated alkyl substances.

Perfluorinated alkyl substances (PFASs) are persistent, toxic, ubiquitously distributed, and bioaccumulated substances, which have attracted increasing concern. To investigate the environmental effects of PFASs, there is a need to develop a sensitive, rapid, and efficient method for detecting trace level PFASs. In this study, a conjugated microporous polymer (CMP) with loading of fluorine, fabricated by Sonogashira-Hagihara cross-coupling, was exploited as a solid-phase extraction (SPE) adsorbent. The prepared fluorine-functionalized CMP (FCMP), which showed a large surface area of 1089 m2·g-1, high porosity, and good chemical stability, was used to extract PFASs from water samples. The adsorption mechanism was investigated using a sorption isotherm model, and the main interactions were fluorous and hydrophobic affinity. The FCMP-based SPE combined with high-performance liquid chromatography-tandem mass spectrometry achieved low limits of detection (0.19-0.97 ng·L-1), wide linear range (2-1600 ng·L-1), and good reproducibility (3.4%-12.9%) under the optimal conditions. Furthermore, the approach was utilized for the analysis of three water samples (snow, river water, and irrigation water) to evaluate its reliability, and satisfactory recovery (70.5%-127.5%) was obtained. Thus, FCMP was feasible SPE adsorbents for the selective extraction of PFASs.

Therapeutic effect of scalp-based acupuncture and moxibustion as an adjunctive treatment on children with cerebral palsy comparing to conventional rehabilitation therapy: a systematic review and meta-analysis of randomized controlled trials.

Background: Cerebral palsy (CP) in children is a predominantly congenital developmental disease with complex causes and diverse symptoms. Chinese medicine mainly uses acupuncture for the treatment of CP; as the disease site is in the brain, emphasis is placed on scalp acupuncture therapy. There were studies about the treatment but different studies had very different results. In this study, we performed a systematic review and meta-analysis of the recent reports on scalp acupuncture in the treatment of CP in children, providing evidence for clinical diagnosis and treatment. Methods: The databases of PubMed, Chinese Biomedical Literature (CBM), China National Knowledge Infrastructure (CNKI), and VIP were searched for randomized controlled trials (RCTs) on scalp acupuncture treatment of pediatric CP published from January 2000 to December 2021. The inclusion criteria of studies were made according to the Participants, Intervention, Control, Outcomes, Study design (PICOS) principles. The Cochrane risk of bias 2.0 was used to evaluate the bias of the included literature. Meta-analysis was performed using the effective rate, Mental Development Index (MDI), Psychological Development Index (PDI), and Gross Motor Function Measure (GMFM-88 scale) as outcome indicators for the efficacy, and the safety of scalp acupuncture was assessed. Results: Initially, 332 articles were retrieved; after screening, 11 articles were included in the selection, including a total of 731 children, with 369 and 362 children for the experimental group and control group respectively. Meta-analysis showed that scalp acupuncture significantly improved the symptoms of children with CP [odds ratio (OR) =3.73, 95% confidence interval (CI): 2.49-5.58, Z=6.41, P<0.00001], could significantly improve their mental development [mean difference (MD) =15.58, 95% CI: 11.74-19.43, Z=7.95, P<0.00001] and psychological development (MD =13.23, 95% CI: 6.17-20.28, Z=3.67, P=0.0002) of children, and significantly improved the motor ability of CP children (MD =17.45, 95% CI: 8.19-26.72, Z=3.69, P=0.0002). Discussion: The curative effect of scalp acupuncture is better than that of conventional rehabilitation. Scalp-based acupuncture therapy can effectively improve the symptoms of pediatric CP, promote the mental and psychological development of children, and improve their gross motor function, the treatment is safe.