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Ovarian cysts are one of the most common gynecological diseases, and laparoscopic surgery has been considered as the main approach for treating benign ovarian tumors. Laparoendoscopic single site surgery (LESS) is favored by women due to its hidden scar in the umbilicus. Combining single-port laparoscopy with open surgery (OPEN) can enhance aesthetic outcomes, reduce surgical complexity, and preserve ovarian function better. The concept of LESS-OPEN pioneered by our team in China has shown significant efficacy in treating ovarian cysts. We also pioneered the construction of a feasibility prediction scoring system for laparoendoscopic single-site extracorporeal (LESS-E) ovarian cystectomy, and established an indication system for LESS-E surgery for ovarian cysts to guide surgeons in selecting surgical approaches. This paper aims to discuss the application of the LESS-OPEN technique in ovarian cystectomy and to explore its future prospects.

Objective: To construct models of high-risk human papillomavirus (HPV) infection with precancerous lesions or cervical cancer and explore the immune function. Methods: Using CRISPR/Cas9, the expression vector HPV16-E6-E7-Rosa26 was microinjected into fertilized eggs of C57BL/6 N mice using homologous recombination, and the F0 generation was obtained for reproduction. Then, the formation of precancerous lesions was promoted via intramuscular injection of estradiol. Presence of precancerous cervical-vaginal intraepithelial lesions, Ki67 and p16 expression levels, and CD8+ T cell proportions in the spleen were evaluated. Results: Two F0 generation mice exhibited correct the homologous recombination. Seven positive mice were identified in the F1 generation. After breeding and mating, 25 homozygous and 11 heterozygous HPV16-E6-E7-engineered mice were obtained from the F2 generation. After estradiol benzoate treatment, the cervical-vaginal epithelium appeared as precancerous lesions with positive Ki67 and p16 expression. The percentage of CD8+ T cells decreased. Conclusion: HPV16-E6-E7-Rosa26 induced low immune function in mice, and provides a good model for the basic research of the mechanisms of action of HPV infection-associated precancerous lesions or cervical cancer.

Objective To investigate the biological and metabolic roles of WNK2 in regulation of ovarian cancer based on transcriptome sequencing technology. Methods Ovarian cancer SKOV3 cells were divided into experimental group and control group, with 3 replicates in each group.The expression of WNK2 was knocked down with transfection of siRNA targeting WNK2 in SKOV3 cells.Then the total RNA was extracted and prepared for transcriptome sequencing and bioinformatics analysis after 48 h of transfection.The differential genes between siNC and siWNK2 were screened out in the range of padj < 0.01, fold change >1, and FPKM >500 to obtain the genes under the regulation of WNK2.DisGeNET and disease ontology (DO) enrichment analysis was used to analyze the relationship of the diseases and differentially expressed genes (DEGs).GO enrichment analysis was adopted to analyze the cellular functions and metabolic processes in which the DEGs were enriched.Finally, RT-qPCR was used to examine whether the 10 differential proteins in the reactive oxygen species (ROS) pathway were regulated by WNK2.The influence of WNK2 overexpression and knockdown on the ROS metabolic process of ovarian cancer cells was determined. Results The DisGeNET and DO enrichment analysis showed the obtained DEGs were closely related with malignant tumors (P < 0.05).GO functional enrichment analysis showed the obtained DEGs were enriched in biological processes such as ROS metabolism, apoptosis signaling pathway, formation of ubiquitin ligase complex and mitochondrial membrane protein complex, growth factor binding, and transcription activator activity and so on (P < 0.05).Among these enriched pathways, ROS related pathways were the most numerous and significant (P < 0.001).The results of RT-qPCR indicated that the changes in 9 DEGs were consistent with the sequencing results after WNK2 was knocked down, and one gene (PDGFB) was contrary to the sequencing results, and one gene had no significant change between the 2 groups.Knockdown of WNK2 in SKOV3 and CAOV3 cells resulted in decreased production of ROS, whereas overexpression of WNK2 increased the production. Conclusion WNK2 probably promotes ovarian cancer malignancy by inducing ROS metabolism.

Objective To investigate the safety of the ultra-early (0~6 h) activity program for patients after minimally invasive radical treatment of cervical cancer. Methods A non-randomized concurrent controlled trial was conducted on 118 patients undergoing minimally invasive radical surgery for cervical cancer in our department from June to December 2019. According to patient's wish, they were divided into the ultra-early (0~6 h) postoperative activity group (n=58) and the conventional activity group (n=60). The patients in both groups received the perioperative program of rapid gynecological rehabilitation. After the operation, the conventional activity group adopted the conventional nursing plan, and the ultra-early activity group adopted the ultra-early postoperative activity plan according to the characteristics of patients with one-to-one guidance and assistance by professional health education nurses. If a patient suffered from conscious weakness, palpitation, dizziness, pale or cold sweats, oxygen saturation < 94%, or heart rate reaching the target heart rate of Karvonen formula, the activity program would be stopped immediately when any of them presented. Results The time to get out of bed for the first time and the activity scores were better in the ultra-early activity group than the conventional activity group (P=0.011). There were no statistically significant differences in SpO2, pulse, pain scores, and incidence of complications between the 2 groups before and after activities. The ultra-early activity group had obviously better average activity in 3 d post-operatively during hospitalization (P=0.000), shorter times to first postoperative exhaust (P=0.020) and to first oral feeding (P=0.010), and shorter length of hospital stay (P=0.010) when compared with the conventional activity group. Conclusion All patients in the ultra-early activity group are able to get out of bed within 24 h post-operatively, and attain better activity level in 3 d after operation than those of the conventional activity group. Ultra-early postoperative activity program can effectively improve the early out-of-bed activity rate after cervical cancer surgery and have no effect on the occurrence of postoperative complications.

Background X-linked hypophosphatemia (XLHR) is the most common genetic form of hypophosphatemic rickets (HR), which is caused by phosphate regulating endopeptidase homolog X-linked (PHEX) gene mutation. At present, the genotype–phenotype relationship of XLHR and the pathogenic role of PHEX are not fully understood. Methods In this study, we summarized clinical features in a new cohort of 49 HR patients and detected 16 novel PHEX and 5 novel non-PHEX variants. Subsequently, we studied the pathogenesis of new variants by protein expression, glycosylation analysis, subcellular localization, and endopeptidase activity. Results The results showed that missense variants (Q189H and X750R) slightly reduced protein expression without obviously altering protein length and localization, whereas truncating variants significantly impaired the synthesis of PHEX and produced a shorter immature protein in cells. Interestingly, no evident correlation was observed between mutation types and clinical phenotypes. However, when we analyzed the relationship between PHEX activity and serum phosphorus level, we found that patients with low PHEX activity tended to have severe hypophosphatemia and high rickets severity score. Following this observation, we established 2 new knock-in XLHR mouse models with 2 novel Phex variants (c.T1349C and c.C426G, respectively) using CRISPR/Cas9 technology. Both mouse models demonstrated clinical manifestations of XLHR seen in patients, and PhexC426G mice showed more severe phenotype than PhexT1349C mice, which further confirmed the rationality of genotype–PHEX enzymatic activity correlation analysis. Conclusion Therefore, our findings demonstrated that novel PHEX variants could disrupt protein function via affecting protein synthesis, post-translational modification, cellular trafficking, and catalytic activity. Our study facilitates a better understanding of XLHR pathogenic mechanism and PHEX activity-phenotype correlation, which is of crucial importance for future diagnosis and treatment of XLHR. [ABSTRACT FROM AUTHOR]

Objective To construct a deep learning automatic segmentation model based on the magnetic resonance image (MRI) of the pelvic floor, and make the intelligent segmentation of the pelvic floor MR image so as to reduce the work intensity of doctors and improve the segmentation efficiency and accuracy of levator ani muscle. Methods Based on DenseUnet model, a network structure composed of encoder module, context extraction module and decoder module was established; In the context extraction module, we used dilated convolution and pyramid pooling module to overcome the disadvantages that Unet uses less context information and global information under different receptive fields. We employed the MRI data of 19 patients, including 14 normal cases, 1 case of grade 1 pelvic organ prolapse (POP1) and 2 cases of grade 2 pelvic organ prolapse (POP2) as training sets. One normal pelvic floor MRI image and 1 POP2 pelvic floor MR image were used for verification. Results The model can segment levator ani muscle in pelvic floor MR image automatically and effectively. Through verification, the average similarity coefficient of levator ani muscle in the test set is 77.1%, the average Hausdorff distance is 16 mm, and the average symmetry plane distance is 0.9 mm. The average similarity coefficient of levator ani muscle in normal volunteers was 81.2%, and that of POP2 female pelvic floor levator ani muscle was 74.5%. Conclusion The segmentation accuracy of DenseUnet model is better than that of Unet, ResUnet and Unet++. It has a strong practical value in the automatic segmentation task of levator ani muscle in MRI images. Through the automatic segmentation of levator ani muscle, the repetitive work of doctors is reduced, and the work efficiency is improved. At the same time, it also provides an alternative for the intelligent auxiliary diagnosis and treatment of pelvic organ prolapse.

Ramie is one of the most important fiber crops in China, with fibers extracted from stem barks having been used as textile materials for thousands of years. DNA methylation is an important epigenetic modification involved in plant growth and development. However, the role of methylation in ramie fiber growth remains poorly understood. In the present study, we investigated the DNA methylation landscape of the nuclear genome in bark sections taken from the top (TPS) and the middle (MPS) of the stems of ramie plants, which represent different stages of fiber growth, using whole-genome bisulfite sequencing. We detected 7,709,555 and 8,508,326 5-methylcytosines in the TPS and MPS genomes, respectively. The distribution of methylation across three sequence contexts, CG, CHG, and CHH, varied greatly among gene elements, with methylation at CHH being the most prevalent. Comparison of methylation levels between the TPS and MPS genomes revealed 23.162 Mb of differentially methylated genomic regions, encompassing 9485 genes. Among these differentially methylated genes, 841 exhibited altered expression in the MPS genome. Notably, an SND2 ortholog Bni05G006779 showed a negative correlation between its expression and methylation levels. Overexpression of Bni05G006779 in Arabidopsis dramatically increased the number of xylem fibers and the secondary wall thickness of the fibers in the stems of transgenic plants. These findings provide important insights into the involvement of DNA methylation in regulating ramie fiber growth. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression of folate receptor 1(FOLR1) and mesothelin (MSLN) in epithelial ovarian cancer(EOC) and explore their clinical significance. Methods The data of clinical samples and transcriptome sequencing expression matrix of EOC were obtained from TCGA and GEO database(Ovarian Cancer TCGA, Provisional; GSE6008; GSE66957; GSE49997) to analyze the expressions of FOLR1 and MSLN in different tissues and their association with tumor staging and the prognosis of EOC. We also collected paraffin-embedded tissue samples of EOC(105 cases), ovarian borderline tumor(11 cases), and normal ovarian tissue(30 cases) from patients undergoing surgeries in our hospital between 2018 and 2019. The expression levels of FOLR1 and MSLN were detected by immunohistochemistry and their correlation with the clinicopathological characteristics of the patients was analyzed. Results Analysis of TCGA and GEO databases showed that the mRNA levels of FOLR1 and MSLN were significantly higher in EOC tissues than in normal ovarian tissues(P < 0.05), and was also higher in stage Ⅲ~Ⅳ than in stage Ⅰ~Ⅱ EOC(P < 0.05). Survival analysis showed that the patients with a high expression of FOLR1 had significantly longer overall survival(P < 0.05) and disease-free survival(P < 0.05), while the patients with a high expression of MSLN had significantly shortened overall survival(P < 0.05) and disease-free survival(P < 0.05). Immunohistochemical analysis showed that FOLR1 and MSLN proteins were lowly expressed in normal ovarian tissues, and their expressions increased moderately in borderline ovarian tumor and significantly in EOC tissues. The positive expression rates of FOLR1 and MSLN in EOC were 80.95% and 53.33%, respectively. The levels were of FOLR1 and MSLN differed significantly in different tissue types(P < 0.001), and their positive rates were significantly higher in stage Ⅲ~Ⅳ than in stage Ⅰ~Ⅱ EOC(P < 0.05). The positive rate of MSLN was significantly increased in patients with lymph node metastasis(P < 0.05). Subgroup survival analysis of TCGA database showed that patients with a low MSLN expression and a high FOLR1 expression(MLFH) at the mRNA level had the best prognosis, while those with a high MSLN expression and a low FOLR1 expression(MHFL) had the worst prognosis(median survival time 70.57 vs 25.85 months, P < 0.000 1); compared with the MLFL patients, the MHFL patients in the early stage(< 50 months) had a better prognosis, while beyond 50 months, the survival rate of the patients decreased, but the difference was not statistically significant. Conclusion In patients with EOC, a high expression of FOLR1 indicates a more favorable prognosis while a high expression of MSLN is associated with a poor prognosis. These 2 proteins may participate in the process of EOC, and their combined detection may help in prognostic evaluation of EOC.