Long-term Follow-up of photodynamic therapy of cervical intraepithelial neoplasia grade 2 (CIN2).

BACKGROUND: To determine the long-term efficacy and safety of 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) for treating cervical intraepithelial neoplasia grade 2 (CIN2) as well as the suitability of ALA-PDT in treating of cervical lesions divided into cervical transformation zone type 3. METHODS: We included 81 patients diagnosed with CIN2 at the Department of Gynecology of the Affiliated Hospital of Qingdao University with data collected between January 2019 and January 2021 following ALA-PDT. Furthermore, we analyzed the superiority of ALA-PDT in fertility preservation among women of childbearing age based on follow-up data from 11 patients with fertility requirements. RESULTS: Our findings confirmed the long-term efficacy of ALA-PDT for CIN2 treatment, with an overall efficacy of 95.83% (23/24) at follow-up of 25-36 months. Moreover, the cervical transformation zone type 3 improvement and human papillomavirus (HPV)-negative efficacy were 69.2% (18/26) and 82.4% (14/17), respectively. ALA-PDT is recommended for consenting patients with cervical transformation zone type 3. Additionally, women without primary infertility could experience natural pregnancy and full-term birth of more than one baby following ALA-PDT for CIN2 treatment, with a satisfaction rate of ≈100%. CONCLUSIONS: ALA-PDT is recommendable for treating high-grade squamous intraepithelial lesions, especially in patients with fertility requirements.

Case report: A case of recurrent cervical cancer with bronchial and esophageal metastases presenting with hemoptysis and dysphagia.

Background: The treatment outcomes and prognosis for recurrent cervical cancer are generally poor, with a 5-year survival rate of only 10%-20%. Case presentation: In this case, the patient is a young woman who experienced a recurrence 5 years after the initial treatment of cervical cancer. Her primary symptoms were hemoptysis and dysphagia, indicative of hilar and mediastinal lymph node metastases, with further involvement of the bronchus and esophagus. Additionally, the patient also presented with tumor-associated dermatomyositis. Following combined treatment with albumin-bound paclitaxel, carboplatin, bevacizumab, and cadonilimab, the patient's tumor was effectively controlled.

Role of extracellular vesicle in human papillomavirus-associated cervical cancer.

BACKGROUND: Cervical cancer is a gynecological malignant tumor and a serious threat to women's health. Although human papillomavirus (HPV) infection and the occurrence of cervical cancer are known to be closely related, the underlying carcinogenic mechanism of HPV is not fully understood. Extracellular vesicles (EVs) are found in a variety of body fluids and play an important role in both intercellular communication and cancer progression. Furthermore, the presence of EVs makes liquid biopsy of cervical cancer possible. The study of EVs in cervical cancer can provide clinical ideas for the diagnosis and treatment of the disease. OBJECTIVES: The purpose of this article is to summarizes the role of EV contents in HPV-associated cervical cancer and discusses the possible clinical application of EVs in cervical cancer treatment. METHODS: The search terms included the following: HPV with cervical cancer and extracellular vesicles. The initial literature search ended on March 1, 2023. CONCLUSIONS: In HPV-positive cervical cancer, EV contents are changed due to the presence of HPV. HPV-positive cervical cancer affects the cell microenvironment and other surrounding cells through the secretion of EVs.

Investigation of periodic characteristics of perturbed flow over a slender body.

The asymmetric flow over a slender body was particularly sensitive to the nose at a high angle of attack (AoA). Two patterns of separation occurred on the noses of the pointed-nosed slender body and blunt-nosed slender body as open- and close-type separation, respectively. The effects of the bluntness were investigated at high AoA (α = 50°) to clarify the evolution of the separated pattern from open-to close-type separation by the nose and by the periodic characteristics of perturbed flow. Wind tunnel experimental tests were conducted to investigate the periodic characteristics of asymmetric flow at a Reynolds number ReD = 1.54 × 105, based on incoming free-stream velocity (U∞) and the diameter (D) of the model. A particle was attached to the tip of the nose to induce the perturbed flow and attain a definite and predictable asymmetric flow in experimental tests. The pressure scanning and surface oil-flow visualization techniques were used to capture the pressure distributions and flow separations. The major findings were that axial flow increases with the increase of bluntness, resulting in open-type separation turning into close-type separation, and the perturbation moved from downstream to upstream of starting points of the separation line. The critical bluntness of separation pattern switching from open-type to close-type located between 1.5 and 3. Thus, the management of perturbation on asymmetric flow pattern switched from directly participating in separation to influencing separation through micro-flow. Therefore, the locations of perturbation and starting points of the separation line were closely related to asymmetric flow management by perturbation, then affecting the periodic characteristics of perturbed flow.

Comparison of the clinical characteristics and prognosis between clear cell carcinomas and high-grade serous ovarian carcinomas.

OBJECTIVES: To compare the clinical characteristics and prognosis of women with clear cell versus high-grade serous ovarian carcinoma. MATERIAL AND METHODS: Retrospective analysis of the clinical data of 50 cases patients with ovarian clear cell carcinoma (OCCC) and 103 cases with high-grade serous ovarian carcinoma (HGSOC), who were initially treated and completed standardized therapy in Affiliated Hospital of Qingdao University from January 2013 to December 2017. RESULTS: There were significant differences in age, gravidity (G > 1), chief complaint, with ovarian endometriosis, tumor diameter, unilateral or bilateral, cystic and solid tumor, CA125, HE4, CA199, lactate dehydrogenase (LDH), and FIGO stage between the two groups. The differences in the prognosis between OCCC patients and HGSOC patients with early stage (FIGO I-II) were not statistically significant. The 5-year overall survival and progression-free survival of OCCC patients were significantly worse than those of HGSOC patients with advanced stage (FIGO III-IV) (p < 0.05). FIGO stage and non-R0 resection were independent risk factors affecting the prognosis of patients with ovarian clear cell carcinoma, screening by Cox regression analysis. FIGO stage, the lowest value of CA125, and non-R0 resection were independent risk factors affecting the prognosis of patients with high-grade serous ovarian cancer. CONCLUSIONS: The clinical characteristics and prognosis of OCCC are different from those of HGSOC. Ovarian clear cell carcinoma (OCCC) patients have a significantly worse prognosis than those with HGSOC in the advanced stage (FIGO Ⅲ-Ⅳ). Satisfactory tumor resection is an essential factor related to the prognosis of patients with OCCC and HGSOC.

Distinct miRNAs associated with various clinical presentations of SARS-CoV-2 infection.

MicroRNAs (miRNAs) have been shown to play important roles in viral infections, but their associations with SARS-CoV-2 infection remain poorly understood. Here, we detected 85 differentially expressed miRNAs (DE-miRNAs) from 2,336 known and 361 novel miRNAs that were identified in 233 plasma samples from 61 healthy controls and 116 patients with COVID-19 using the high-throughput sequencing and computational analysis. These DE-miRNAs were associated with SASR-CoV-2 infection, disease severity, and viral persistence in the patients with COVID-19, respectively. Gene ontology and KEGG pathway analyses of the DE-miRNAs revealed their connections to viral infections, immune responses, and lung diseases. Finally, we established a machine learning model using the DE-miRNAs between various groups for classification of COVID-19 cases with different clinical presentations. Our findings may help understand the contribution of miRNAs to the pathogenesis of COVID-19 and identify potential biomarkers and molecular targets for diagnosis and treatment of SARS-CoV-2 infection.

Clinical outcome of photodynamic therapy for the treatment of cervical intraepithelial neoplasia grade 2.

Five-aminolaevulinic acid photodynamic therapy (ALA-PDT) was used to treat 79 cervical intraepithelial neoplasia 2 (CIN 2) patients who desired preservation of fertility ,between Oct 2018 and Dec 2020. Three months after treatment, among the 65 patients who returned for follow-up, full recovery and improvement rates were 43/65 and 16/65, respectively, resulting in a total response rate of 90.77%. This suggests that ALA-PDT is worthy of clinical application, even as monotherapy. The result of immune testing also indicated significant promotion of CD4+T expression during the recovery process, highlighting the importance of immune responses in different prognoses.

Identification of Distinct Immune Cell Subsets Associated With Asymptomatic Infection, Disease Severity, and Viral Persistence in COVID-19 Patients.

The cell-mediated protective and pathogenic immune responses to SARS-CoV-2 infection remain largely elusive. Here we identified 76 distinct cell subsets in the PBMC samples that were associated with various clinical presentations of COVID-19 using scRNA-seq technology coupled with a deep and comprehensive analysis of unique cell surface markers and differentially expressed genes. We revealed that (TRAV1-2+CD8+)MAIT cells and (NCAM1hiCD160+)NK cells significantly enriched in the asymptomatic subjects whereas (LAG3+CD160+CD8+)NKT cells increased in the symptomatic patients. We also observed that (CD68-CSF1R-IL1BhiCD14+)classical monocytes were positively correlated with the disease severity. Moreover, (CD33-HLA-DMA-CD14+)classical monocytes and (CLEC10A-S100A9lo)pDC were associated with the viral persistence. The GO and KEGG analyses identified enriched pathways related to immune responses, inflammation, and apoptosis. These findings may enhance our understanding of the immunopathogenesis of COVID-19 and help develop novel strategies against SARS-CoV-2 infection.

T-box transcription factor TBX1, targeted by microRNA-6727-5p, inhibits cell growth and enhances cisplatin chemosensitivity of cervical cancer cells through AKT and MAPK pathways.

Cervical cancer (CC) is the fourth most common cancers among women worldwide. T-box transcription factor 1 (TBX1), a member of the T-box family, has anti-tumor effects in some types of cancer, but its role in CC is yet unknown. The aim of this study is to investigate the functions and underlying mechanisms of TBX1 in CC. Online database UALCAN showed that TBX1 was down-regulated in CC tissues compared with normal tissues and patients with lower TBX1 expression level had a poor prognosis. TBX1 overexpression significantly decreased the proliferation, migration, and invasion of Hela and SiHa cells. Conversely, cell apoptosis and chemosensitivity to cisplatin were promoted in TBX1-overexpressing CC cells. Moreover, up-regulation of TBX1 inhibited both AKT and MAPK signaling pathways. Furthermore, dual luciferase report assay indicated that TBX1 could directly bind to miR-6727-5p. In addition, TBX1 expression was inhibited by miR-6727-5p mimic and up-regulated by miR-6727-5p inhibitor. Knockdown of TBX1 reversed the inhibitory effect of the miR-6727-5p inhibitor on CC cells. This study demonstrates that TBX1, a target gene of miR-6727-5p, acts as a tumor suppressor in CC, indicating that TBX1 may be a new target for CC therapy.

RHPN2 Promotes Malignant Cell Behaviours in Ovarian Cancer by Activating STAT3 Signalling.

BACKGROUND: Ovarian cancer is the fifth most common cause of cancer-related deaths and accounts for 3% of cancer cases occurring in women. Therefore, determining the underlying genes that can promote ovarian cancer progression is of great urgency. It has been reported that RHPN2 promotes tumour progression in various types of cancer, but its role in ovarian cancer pathogenesis remains unknown. MATERIALS AND METHODS: In this study, bioinformatic datasets were used to predict the expression of RHPN2 in clinical samples and determine the relationship between RHPN2 and the prognosis of ovarian cancer patients. Clinical samples were used to verify the prediction. RHPN2-targeting shRNA was used to investigate the effect of RHPN2 on ovarian cancer cells, and following RHPN2 knockdown, the proliferative and migratory capacities of ovarian cancer cells were tested. To determine the downstream signalling target of RHPN2, a luciferase reporter assay was conducted, and an animal experiment was carried out to confirm the effect of RHPN2 in vivo. RESULTS: The public datasets indicated that ovarian cancer tissues showed significantly higher RHPN2 expression than para-cancer normal tissues, and poor prognosis was observed in patients with higher RHPN2 expression, which was further confirmed in clinical samples. After RHPN2 was knocked down, the proliferation and migration of ovarian cancer cells were significantly impaired; a luciferase reporter assay indicated that the STAT3 signalling pathway was the most highly affected, and RHPN2 downregulation inhibited STAT3 nuclear translocation. STAT3 inhibitors partially rescued the tumour-promoting effect induced by RHPN2 overexpression, which was further confirmed by animal experiments. CONCLUSION: Collectively, our results indicate that RHPN2 promotes malignant behaviours in ovarian cancer by activating STAT3 signalling.

Loop electrosurgical excision procedure followed by 5-aminolevulinic acid photodynamic therapy for cervical intraepithelial neoplasia, a report of six cases.

BACKGROUND: Cervical intraepithelial neoplasia (CIN) may progress to cervical cancer if left untreated. The loop electrosurgical excision procedure (LEEP) of the transitional zone of the cervix is a standard form of treatment. However, human papillomavirus (HPV)-induced CIN may recur after LEEP. The purpose of this case report is to describe the successful use of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) as an adjunct to LEEP, in preventing the recurrence of CIN. METHODS: The effectiveness of this combined treatment was evaluated in six women. The diagnosis of HPV-induced CIN was determined using HPV DNA tests and liquid-based cervical cytology. Lesion removal was performed 3 h after application of ALA using a 635 nm light density of 80 mw/cm2. RESULTS: We treated 6 women aged 31-62 years who had persistent CIN following LEEP, with ALA-PDT (range, 4-7 treatments). Five of the 6 women were HPV negative on retesting 6-7 months after ALA-PDT. Most patients showed no signs of recurrence during the follow-up period. CONCLUSIONS: Use of ALA-PDT following LEEP may prevent the recurrence of CIN. Monitoring HPV status by means of DNA testing and liquid-based cytology may be used as a standard for clinical diagnosis and treatment. Post-treatment care should be carefully considered because improper post-treatment care might directly lead to relapse.

C-Myc-activated long non-coding RNA PVT1 enhances the proliferation of cervical cancer cells by sponging miR-486-3p.

Cervical cancer is one of the most prevalent gynecological malignancies. Although the functions of long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) and c-Myc in tumorigenesis have been acknowledged, the roles of c-Myc and lncRNA-PVT1 in the proliferation of cervical cancer are still unclear. Our study is designed to demonstrate the regulatory network involving c-Myc and lncRNA-PVT1 in cervical cancer. Quantitative real-time PCR and western blot assays were performed in our research to estimate the expression levels of RNA and proteins. CCK8 assays were applied to demonstrate the viability of HeLa and SiHa cells. Immunofluorescence assay was then used to investigate the co-localization of lncRNA-PVT1 and miR-486-3p. Binding of c-Myc to the promoter region of PVT1 was identified by ChIP-assay. Functionally, upregulation of lncRNA-PVT1 enhanced the proliferation and viability of cervical cancer cells. Mechanistically, lncRNA-PVT1 sponged miR-486-3p and released its repression of extracellular matrix protein 1. Besides, c-Myc functioned as an activator of lncRNA-PVT1 and upregulated its expression by binding to the promoter of PVT1 in cervical cancer cells. lncRNA-PVT1 was upregulated by c-Myc and thus enhanced the proliferation of cervical cancer cells by sponging miR-486-3p.

Long non­coding RNA plasmacytoma variant translocation 1 gene promotes the development of cervical cancer via the NF­κB pathway.

The long noncoding RNA plasmacytoma variant translocation 1 gene (LncRNA PVT1) has an important role in tumor occurrence and development, yet the role and underlying molecular mechanisms of this RNA in cervical cancer have not yet been elucidated. In the present study, three cervical cancer cell lines (HeLa, Ca Ski and SiHa) were used to verify how LncRNA PVT1 mediates cervical cancer development, and the H8 cell line was used as a control. A LncRNA PVT1 overexpression vector or small interfering RNAs targeting LncRNA PVT1 were transfected into cervical cancer cells to generate LncRNA PVT1 overexpression and silencing in these cells. LncRNA PVT1 overexpression accelerated the growth of cervical cancer cells by advancing the cell cycle and inhibiting cellular apoptosis; increases in Cyclin D1 (CCND1) mRNA and activated Bcl­2 protein expression levels also supported this finding. Furthermore, NF­κB activation and expression was increased by LncRNA PVT1 overexpression. In addition, NF­κB activation or inhibition induced changes in cell viability, accompanied by changes in CCND1 and Bcl­2 expression. Increases or decreases in microRNA­16 (miR­16) expression (using miR mimics and inhibitors) also corresponded to changes in LncRNA PVT1 expression, in vitro. miR­16 mimics and inhibitor had opposite effects to those of NF­κB activity, and miR­16 was demonstrated to directly interact with the NF­κB gene as measured using the dual­luciferase assay. In summary, LncRNA PVT1 inhibits the effect of miR­16, promoting the cell cycle and inhibiting cellular apoptosis of cervical cancer cells, potentially via the NF­κB pathway. The data from the present study will contribute to the current knowledge surrounding the theoretical basis of cervical cancer and provide a new perspective for the treatment of cervical cancer.

Intra-tumor heterogeneity for endometrial cancer and its clinical significance.

BACKGROUND: Management of tumors has become more complex owing to tumor heterogeneity. Fewer studies have been performed on intra-tumor heterogeneity of endometrial cancer (EC) until now. Therefore, it is of great clinical value to explore the intra-tumor heterogeneity of EC based on clinical features and gene expression profiles. METHODS: A total of 1688 patients with EC were screened and 114 patients were finally selected, including specimens from 84 patients with primary EC without relapse (PE) and the paired metastases (P-M) specimens, as well as specimens from 30 patients with primary EC with relapse (RPE) and the paired relapsed EC (P-RE) specimens. Microarray and RNA-seq were used to detect gene expression of EC samples. Clinicopathological characteristics and molecular data were compared between PE and P-M groups and between RPE and P-RE groups to explore the intra-tumor heterogeneity of EC. RESULTS: The clinical intra-tumor spatial heterogeneity of pathological type, grade, ER status, and PR status between PE and P-M were 17.9%, 13.1%, 28.6%, and 28.6%, respectively. The clinical intra-tumor spatiotemporal heterogeneity of pathological type, grade, ER status, and PR status between RPE and P-RE were 16.7%, 33.3%, 25.0%, and 37.5%, respectively. Cluster analysis sorts EC samples based on progression type of lesion and their pathological type. There were differentially expressed genes between PE and P-M and between RPE and P-RE, of which gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis were mainly enriched in cell proliferation, the p53 signaling pathway, etc. CONCLUSIONS:: Clinical and molecular data showed that there was spatiotemporal heterogeneity in intra-tumor of EC, which may add to the complexity of diagnosis and therapeutics for EC. Considering the intra-tumor heterogeneity, sequential chemotherapy and precision medicine may be a more suitable treatment plan for EC.

Songorine suppresses cell growth and metastasis in epithelial ovarian cancer via the Bcl­2/Bax and GSK3ß/ß­catenin signaling pathways.

Epithelial ovarian cancer (EOC) is the most frequent cause of cancer­associated mortality among all types of gynecological cancer. The high recurrence rate and the poor 5­year survival rate indicate that more effective therapeutic strategies are required. The aim of the present study was to investigate the role and potential mechanisms of songorine in treating EOC. EOC cells were cultured with different concentrations of songorine, following which MTT and flow cytometric analyses were conducted to measure cell viability and apoptosis. Wound healing and Transwell assays were used to detect cell migration and invasion abilities. Furthermore, associated molecules in the glycogen synthase kinase (GSK)­3ß/ß­catenin and B­cell lymphoma 2 (Bcl­2)/Bcl­2­associated X (Bax) signaling pathways were semi­quantified by western blotting. Finally, tumor size measurements, pathological observations, western blot analysis and toxicological evaluations were performed in SKOV­3 tumor­bearing BALB/c nude mice to investigate the efficacy and safety of songorine. As expected, songorine inhibited EOC cell survival, invasion and migration, promoted EOC cell apoptosis and suppressed mammalian EOC tumorigenic behavior. In particular, GSK3ß inhibitor treatment restored the songorine­induced regulation of the GSK3ß/ß­catenin signaling pathway. Furthermore, in the in vitro and in vivo experiments, songorine consistently downregulated the expression of N­cadherin, vimentin, matrix metalloproteinase (MMP)­2, MMP­9, phosphorylated­GSK3ß, ß­catenin and Bcl­2, and upregulated the expression of E­cadherin, cleaved caspase­3, cleaved caspase­9 and Bax. In conclusion, songorine exerted its anticancer effect through the GSK3ß/ß­catenin and Bcl­2/Bax signaling pathways. These results highlight the potential use of songorine as a novel therapeutic agent for EOC.

Mechanism of ultrasonic impregnation on porosity of activated carbons in non-cavitation and cavitation regimes.

Ultrasonic impregnation has proven to be an effective method to improve surface area and pore volume during preparation of activated carbons. However, the mechanism by which the promotion effect of ultrasonic impregnation is still ambiguous. Fundamental wave pressure (FWP) and broadband integrated pressure (BIP) were used to estimate the non-cavitation (vibration) energy and cavitation energy, respectively. The effects of FWP and BIP on the pore volume, surface area, surface functional groups, and microcosmic morphology were investigated in non-cavitation and cavitation regimes. Ultrasonic vibration promoted the surface enlargement and pore development of activated carbons, and it mainly affected the development of mesopore volume (Vmes) in both the pore volume and the mesopore-size-distribution range. The Vmes was enhanced by 60%-100% in the non-cavitation regime. Ultrasonic cavitation also facilitated porosity development of activated carbons, and it mainly affected the development of specific surface area (SBET) and micropore volume (Vmic). The excessive cavitation led to a decrease of the porosity of activated carbons, so the BIP should be optimized during impregnation. The highest SBET, Vmic, and Vmes for activated carbons were obtained by in the presence of both FWP and BIP, which were enhanced by 29.05%, 30.23%, and 113.33%, respectively, compared with the corresponding value for the activated carbon prepared without using ultrasonic impregnation. This work provided new insight into the role of the acoustic energy present during impregnation in tuning properties of activated carbons.

LncRNA XLOC_006390 facilitates cervical cancer tumorigenesis and metastasis as a ceRNA against miR-331-3p and miR-338-3p.

OBJECTIVE: Cervical cancer is one of the most common malignant tumors. Our previous results showed that long non-coding RNA (lncRNA) XLOC_006390 plays an important role in cervical cancer. In this study, we have explored the mechanism of action of lncRNA XLOC_006390. METHODS: LncRNA XLOC_006390 was proposed to exercise its function as a competing endogenous RNA (ceRNA), and its potential targeted miRNAs was predicted through the database LncBase Predicted v.2. Two miRNAs, miR-331-3p, and miR-338-3p, were chosen for the study. Expression of miRNAs and lncRNA in cervical cancer cells and tissues was detected by reverse transcription polymerase chain reaction. To determine the correlation, silencing of XLOC_006390, over-expression of miR-331-3p, and miR-338-3p was performed in SiHa and Caski cell lines, respectively. RESULTS: Based on the interactive effect between miRNA and lncRNA, miR-331-3p and miR-338-3p were significantly downregulated in cervical cancer cells and tissues, and their expression levels were negatively related to that of lncRNA. Our results also showed that the expression of miR-331-3p target gene NRP2, miR-338-3p target genes PKM2, EYA2 was significantly downregulated when the XLOC_006390 was knocked down. Further, XLOC_006390 was found to facilitate cervical cancer tumorigenesis and metastasis by downregulating miR-331-3p and miR-338-3p expression. CONCLUSION: Taken together, our study demonstrated that XLOC_006390 may serve as a ceRNA and reversely regulates the expression of miR-331-3p and miR-338-3p, thus facilitating cervical cancer tumorigenesis and metastasis.

Reduced nucleic acid methylation impairs meiotic maturation and developmental potency of pig oocytes.

Oocyte meiosis is a complex process coordinated by multiple endocrinal and molecular circuits. Recently, N6-methyladenosine (m6A) epigenetic modification on RNA is revealed to be important for meiotic maturation. However, the molecular mechanism of how m6A modification exerts its effect on oocyte maturation is largely unknown. Here, we showed that endogenous m6A writers (Mettl3 and Wtap) and eraser (Fto) elevated their transcript levels during meiotic maturation of pig oocytes. From germinal vesicle (GV) to metaphase II (MII) stages, global m6A level significantly increased, and existed mostly in ooplasm. Methyl donor (betaine, 16 mM) treatment of porcine cumulus-oocyte complexes (COCs) during in vitro maturation (IVM) significantly boosted nucleic acid m6A level within oocytes, but unchanged meiotic process and oocyte subsequent development. By contrast, methylation inhibitor (cycloleucine, 20 mM) reduced nucleic acid m6A level, and significantly decreased the germinal vesicle breakdown (GVBD) rate, the extrusion rate of the first polar body, and the cleavage and blastocyst rates of parthenotes. In addition, in cycloleucine-treated oocytes Wtap increased but Lin28 decreased their abundances significantly, along with the higher incidence of spindle defects and chromosome misalignment. Furthermore, pT161-CDK1 protein level in pig oocytes was confirmed to be decreased after cycloleucine treatment for 24 h. Taken together, chemical induced reduction of nucleic acid m6A methylation during pig oocyte meiosis could impair meiotic maturation and subsequent development potency, possibly through down-regulating pluripotency marker Lin28 mRNA abundance and disturbing MPF-regulated chromosome/spindle organization.

Association of Dose and Frequency on the Survival of Patients on Maintenance of Hemodialysis in China: A Kaplan-Meier and Cox-Proportional Hazard Model Analysis.

BACKGROUND Dialysis frequency and dose are controversial prognostic factors of hemodialysis morbidity and mortality. The aim of this study was to find out the effect of frequency and dosage of dialysis on mortality and survival in a group of Chinese hemodialysis patients. MATERIAL AND METHODS In total, 183 patients seen from February 2008 to January 2018, who were on maintenance hemodialysis for at least 3 months, were included in the study cohort. An anonymized database of age, gender, diabetic status, comorbidities, date of initiation of dialysis, hematological characters, biochemical variables, and status of survived or died was established from DICOM (Digital Imaging and Communications in Medicine) files of patients. Kaplan-Meier and Cox-proportional hazard model was used for calculation of survival over time at 95% confidence level. RESULTS Overall, the 10-year survival rate was 27%. Kaplan-Meier analysis showed patient survival as 94% at one-year, 59% at 5-years, and 27% at 10-years. Hemoglobin, serum albumin, calcium, potassium, phosphorous, calcium-phosphorous-products, and hemodialysis frequency and the dose had a significant effect on survival. Cox regression proportional hazard model showed that patients with serum albumin level of >4 g/dL were better associated with survival. Patients who underwent twice-weekly hemodialysis had 4.26 times less chance of survival as compared to patients with thrice-weekly hemodialysis. A higher dialysis dose of >1.2 spKt/V offered better survival as compared to a lower dose of <1.2 spKt/V. CONCLUSIONS Hypoalbuminemia, hemodialysis time, and hemodialysis frequency were significantly associated with mortality.

Heat shock protein 90α couples with the MAPK-signaling pathway to determine meiotic maturation of porcine oocytes.

Heat shock protein 90 (Hsp90) functions as a molecular chaperone in its interaction with clients to influence multiple cellular and physiological processes. However, our current understanding on Hsp90's relationship with mammalian oocyte maturation is still very limited. Here, we aimed to investigate Hsp90's effect on pig oocyte meiotic maturation. Endogenous Hsp90α was constantly expressed at both mRNA and protein levels in porcine maturing oocytes. Addition of 2 µM 17-allylamino-17-demethoxygeldanamycin (17-AAG), the Hsp90 inhibitor, to in vitro mature cumulus-oocyte complexes (COC) significantly decreased Hsp90α protein level (P < 0.05), delayed germinal vesicle breakdown (GVBD) (P < 0.05), and impeded the first polar body (PB1) extrusion (P < 0.01) of porcine oocytes. 2 µM 17-AAG treatment during in vitro maturation also decreased the subsequent development competence as indicated by the lower cleavage (P < 0.001) and higher fragmentation (P < 0.001) rates of parthenotes, whereas no effects on the percentage and average cell number of blastocysts were found. Immunodepletion of Hsp90α by antibody microinjection into porcine oocytes at germinal vesicle and metaphase II stages induced similar defects of meiotic maturation and parthenote development, to that resulted from 2 µM inhibitor 17-AAG. For oocytes treated by 2 µM 17-AAG, the cytoplasm and membrane actin levels were weakened (P < 0.01), and the spindle assembly was disturbed (P < 0.05), due to decreased p-ERK1/2 level (P < 0.05). However, the mitochondrial function and early apoptosis were not affected, as demonstrated by rhodamine 123 staining and Annexin V assays. Our findings indicate that Hsp90α can couple with mitogen-activated protein kinase to regulate cytoskeletal structure and orchestrate meiotic maturation of porcine oocytes.