Complete mitochondrial genomes of Thyreophagus entomophagus and Acarus siro (Sarcoptiformes: Astigmatina) provide insight into mitogenome features, evolution, and phylogeny among Acaroidea mites.

Mites from the Acaroidea (Sarcoptiformes: Astigmatina) are important pests of various stored products, posing potential threats to preserved foods. In addition, mites can cause allergic diseases. Complete mitochondrial genomes (mitogenomes) are valuable resources for different research fields, including comparative genomics, molecular evolutionary analysis, and phylogenetic inference. We sequenced and annotated the complete mitogenomes of Thyreophagus entomophagus and Acarus siro. A comparative analysis was made between mitogenomic sequences from 10 species representing nine genera within Acaroidea. The mitogenomes of T. entomophagus and A. siro contained 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs), and one control region. In Acaroidea species, mitogenomes have highly conserved gene size and order, and codon usage. Among Acaroidea mites, most PCGs were found to be under purifying selection, implying that most PCGs might have evolved slowly. Our findings showed that nad4 evolved most rapidly, whereas cox1 and cox3 evolved most slowly. The evolutionary rates of Acaroidea vary considerably across families. In addition, selection analyses were also performed in 23 astigmatid mite species, and the evolutionary rate of the same genes in different superfamilies exhibited large differences. Phylogenetic results are mostly consistent with those identified by previous phylogenetic studies on astigmatid mites. The monophyly of Acaroidea was rejected, and the Suidasiidae and Lardoglyphidae appeared to deviate from the Acaroidea branch. Our research proposed a review of the current Acaroidea classification system.

Exploration of a novel prognostic risk signatures and immune checkpoint molecules in endometrial carcinoma microenvironment.

In this study, we devoted to investigate immune-related genes and tumor microenvironment (TME) in EC based on The Cancer Genome Atlas (TCGA) database. In total 799 up-regulated and 139 down-regulated immune-related and differentially expressed genes in EC were investigated for functional annotations and prognosis. By a conjoint Cox regression analysis, we built two risk models for OS and DFS, as well as the consistent nomograms. Immune-related pathways were revealed mostly enriched in the low-risk group. By further analyzing TME based on the risk signatures, the higher immune cell infiltration and activation, lower tumor purity and higher tumor mutational burden were found in low-risk group, which presented a better prognosis. Both the expression and immunophenoscore of immune checkpoints PD-1, CTLA4, PD-L1 and PD-L2 increased significantly in low-risk group. These findings may provide new ideas for novel biomarkers and immunotherapy targets in EC.

RNA demethylase ALKBH5 promotes ovarian carcinogenesis in a simulated tumour microenvironment through stimulating NF-κB pathway.

Methylation is the main form of RNA modification. N6-methyladenine (m6A) regulates the splicing and translation of mRNA. Alk B homologue 5 (ALKBH5) participates in the biological regulation of various cancers. However, its role in ovarian carcinogenesis has not been unveiled. In the present study, ALKBH5 showed higher expression in ovarian cancer tissue than in normal ovarian tissue, but lower expression in ovarian cancer cell lines than in normal ovarian cell lines. Interestingly, Toll-like receptor (TLR4), a molecular functioning in tumour microenvironment (TME), demonstrated the same expression trend. To investigate the effect of abnormal TME on ovarian carcinogenesis, we established an in vitro model in which macrophages and ovarian cancer cells were co-cultured. In the ovarian cancer cells co-cultured with M2 macrophages, the expression of ALKBH5 and TLR4 increased. We also verified that TLR4 up-regulated ALKBH5 expression via activating NF-κB pathway. Depending on transcriptome sequencing, m6A-Seq and m6A MeRIP, we found that NANOG served as a target in ALKBH5-mediated m6A modification. NANOG expression increased after mRNA demethylation, consequently enhancing the aggressiveness of ovarian cancer cells. In conclusion, highly expressed TLR4 activated NF-κB pathway, up-regulated ALKBH5 expression and increased m6A level and NANOG expression, all contributing to ovarian carcinogenesis. Our study revealed the role of m6A in ovarian carcinogenesis, providing a clue for inventing new target therapy.

Identification of EPHX2 and RMI2 as two novel key genes in cervical squamous cell carcinoma by an integrated bioinformatic analysis.

Cervical cancer is the fourth most common malignancy in women worldwide and cervical squamous cell carcinoma (CESC) is the most common histological type of cervical cancer. The dysregulation of genes plays a significant role in cancer. In the present study, we screened out differentially expressed genes (DEGs) of CESC in the GSE63514 data set from the Gene Expression Omnibus database. An integrated bioinformatics analysis was used to select hub genes, as well as to investigate their related prognostic signature, functional annotation, methylation mechanism, and candidate molecular drugs. As a result, a total of 1907 DEGs were identified (944 were upregulated and 963 were downregulated). In the protein-protein interaction network, three hub modules and 30 hub genes were identified. And two hub modules and 116 hub genes were screened out from four CESC-related modules by the weighted gene coexpression network analysis. The gene ontology term enrichment analysis and Kyoto encyclopedia of genes and genomes pathway analysis were performed to better understand functions and pathways. Genes with a significant prognostic value were found by prognostic signature analysis. And there were five genes (EPHX2, CHAF1B, KIAA1524, CDC45, and RMI2) identified as significant CESC-associated genes after expression validation and survival analysis. Among them, EPHX2 and RMI2 were noted as two novel key genes for the CESC-associated methylation and expression. In addition, four candidate small molecule drugs for CESC (camptothecin, resveratrol, vorinostat, and trichostatin A) were defined. Further studies are required to explore these significant CESC-associated genes for their potentiality in diagnosis, prognosis, and targeted therapy.

Competing endogenous RNA network of endometrial carcinoma: A comprehensive analysis.

Competing endogenous RNA (ceRNA) network is dysregulated in the initiation and progression of tumors. In the present study, we explored the regulatory mechanism of ceRNA in endometrial carcinoma (EC) and the potential key molecules with potential value in the diagnosis, treatment, and prognosis of EC. The long noncoding RNAs (lncRNAs) and messenger RNAs (mRNAs) expression profiles (552 EC tissues and 35 nontumor tissues) and microRNAs (miRNAs) expression profiles (546 EC tissues and 33 nontumor tissues) were downloaded from The Cancer Genome Atlas database to identify differentially expressed RNAs (DERNAs) in EC. An integrated bioinformatics analysis was used to construct an EC-specific ceRNA network and select key molecules. As a result, 96 differentially expressed lncRNAs (DElncRNAs), 29 differentially expressed miRNAs (DEmiRNAs), and 77 differentially expressed mRNAs (DEmRNAs) were identified. An EC-specific ceRNA network was built based on nine DElncRNAs significantly associated with overall survival. CCNB1 was found as a key gene in EC through the weighted gene coexpression network analysis and protein-protein interaction network analysis. Our ceRNA network showed C2orf48 and LINC00483 might upregulate CCNB1 via competing with miR-183. In addition, we found a subnetwork which contained survival-associated DERNAs (AC110491.1, LINC00483-miR-192-GRHL1). The results of reverse transcription quantitative polymerase chain reaction supported the relative expressions of C2orf48, LINC00483 were upregulated and that of AC110491.1 was downregulated in EC. We further found C2orf48 was upregulated in serous EC, endometrioid EC, and mixed serous and endometrioid EC. LINC00483 was upregulated in mixed serous and endometrioid EC compared with that in the normal tissues according to UALCAN database. In addition, candidate small molecular drugs were screened out by ConnectivityMap based on the 77 DEmRNAs in the ceRNA network. Eventually, C2orf48, LINC00483, and AC110491.1 were identified as three key lncRNAs in EC.

Eleven genes associated with progression and prognosis of endometrial cancer (EC) identified by comprehensive bioinformatics analysis.

BACKGROUND: Endometrial cancer (EC) is one of the female malignant tumors. Endometrial cancer predominately affects post-menopausal women. Bioinformatics analysis has been widely applied to screen and analyze genes in linkage to various types of cancer progression. METHODS: Download the gene expression profile from Gene Expression Omnibus (GEO). Calculate raw expression data according to pre-processing procedures. We performed the "limma" R language package to screen DEGs between Endometrial cancer tissue samples and normal uterus tissue samples. Enrichment of the functions and pathways was analyzed by using clusterprofiler. We utilized Search Tool for the Retrieval of Interacting Genes Database (STRING) to assess protein-protein interaction (PPI) information, and then we used plug-in Molecular Complex Detection (MCODE) to screen hub modules of PPI network in Cytoscape. We also performed functional analysis on the genes in the hub module by using clusterprofiler. Next, we utilized the "WGCNA" package in R to establish co-expression network for the DEGs. The Venn diagram was performed to overlap the gene in key module and hub PPI cluster. We validated the key genes in TCGA, GEPIA, UALCAN and Immunohistochemistry staining obtained from The Human Protein Atlas database. And then we did ROC curve analysis by SPSS. Gene set enrichment analysis (GSEA) and mutation analysis were also performed for hub genes. RESULTS: Functional and pathway enrichment analysis demonstrated that the upregulated differentially expressed genes (DEGs) were significantly enriched in CXCR chemokine receptor binding, chemokine activity, chemokine receptor binding, G-protein coupled receptor binding, RAGE receptor binding, cytokine activity, microtubule binding, receptor regulator activity and microtubule motor activity, and the down-regulated genes were highly enriched in collagen binding. After using STRING software to construct PPI network, 30 prominent proteins were identified and the first two significant modules were selected. In co-expression network, 5 EC-related modules were identified. Among them, the turquoise module has the highest correlation with the EC. We further analyzed the genes in the PPI and turquoise module, and selected eleven key genes related to EC after validation of TCGA database, GEPIA, UALCAN and immunohistochemistry. Six of them had mutation significance. CONCLUSIONS: In summary, these 11 genes may become new therapy targets for EC treatment.

Tumor-suppressor role of miR-139-5p in endometrial cancer.

BACKGROUND: Endometrial cancer (EC) is the fourth most common malignancy of the female genital tract worldwide. MicroRNAs are important gene regulators with critical roles in diverse biological processes, including tumorigenesis. Several study's show that miR-139-5p is involved in the tumorigenesis and metastasis of various cancers. However, its expression and potential biologic role in endometrial cancer remain to be determined. This study aimed to investigate the miR-139-5p expression and to analyze its function and underlying molecular mechanism in endometrial cancer. METHODS: Expression of miR-139-5p was measured using qRT-PCR. The expression of HOXA10 was detected by Immunofluorescence staining in endometrial cancer tissues and adjacent normal tissues. CCK-8 and colony formation assays were used to assess the effect of miR-139-5p on ECC1 and Ishikawa cell line proliferation. Transwell migration assay was used to study the effect of miR-139-5p on EC cell migration. Luciferase reporter assay and western blot were used to confirm targeting of HOXA10 by miR-139-5p. RESULT: We demonstrated that miR-139-5p was down-regulated in human endometrial cancer compared to their matched adjacent non-tumor tissues. Overexpressed miR-139-5p significantly inhibited endometrial cancer cell viability and migration. Computational algorithm in combination with dual luciferase reporter assays identified HOXA10 as the target of miR-139-5p. HOXA10 expression was downregulated in endometrial cancer cells after miR-139-5p overexpression. The expression level of HOXA10 was significantly increased in endometrial cancer tissues, which was inversely correlated with miR-139-5p expression in clinical endometrial cancer tissues. CONCLUSION: These findings indicate that miR-139-5p targets the HOXA10 transcript and suppresses endometrial cancer cell growth and migration, suggesting that miR-139-5p acts as a tumor suppressive role in human endometrial cancer pathogenesis.

Glucose-Related Protein 78 Expression and Its Effects on Cisplatin-Resistance in Cervical Cancer.

BACKGROUND GRP78, the 78-kDa glucose-regulated protein, occupies a significant position in endoplasmic reticulum stress. Emerging evidences have shown that GRP78 induces chemoresistance in several tumors; however, the role of GRP78 in cervical cancer (CVC) still needs to be elucidated clearly. MATERIAL AND METHODS In the present study, we determined the expression levels of GRP78 in CVC tissues collected from patients through immuocytochemistry, western blot and real-time PCR. To evaluate the exact role of GRP78 in CVC cells in the presence of cisplatin, we generated GRP78 knock-down cervical cancer cells through small interfering RNA. After successful transfection, the apoptosis rate was assessed with flow cytometry. The expression levels of caspase-3, CHOP and Bcl-2 in GRP78 knock-down cells were determined by western blot. RESULTS The GRP78 levels in CVC tissues were increased significantly. Three types of CVC cells HeLa, SiHa, and C33A were treated with different concentrations of cisplatin and cultured for 12 hours, 24 hours, and 48 hours respectively. And SiHa cells exhibited the highest resistance to cisplatin at all time. Specifically, after 25 µM cisplatin treatment, more than 80% of C33A cells underwent apoptosis, whereas the apoptotic rate of SiHa cells was only 30-40%. Data suggested that GRP78 silencing increased chemo-sensitivity and improved the effects of cisplatin-induced apoptosis in SiHa cells. Moreover, inhibition of GRP78 could upregulate caspase-3 and CHOP expression and downregulate Bcl-2 expression. CONCLUSIONS GRP78 may represent a key bio-marker of CVC and silencing GRP78 may strengthen the resistance against cisplatin. GRP78 may be a potential molecular target for CVC therapies in future.

A Polysaccharide from Amusium Pleuronectes Combined with Praziquantel Treatment Ameliorates Hepatic Fibrosis in Schistosoma Japonicum-Infected Mice.

BACKGROUND Polysaccharides from bivalves have multiple bioactivities in various aspects of biology. However, the role of a polysaccharide derived from Amusium pleuronectes on potential hepatoprotective effects remains unclear. MATERIAL AND METHODS A water-soluble polysaccharide was isolated from Amusium pleuronectes (APS-1) using ultrasound-assisted hot-water extraction. The molecular weight of APS-1 was approximately 11.7 kDa and was determined by calibration with dextran. APS-1 was analyzed by high-performance liquid chromatography (HPLC), and mainly consisted of a uniform glucose polymer. The protective effect of APS-1 on Schistosoma japonicum-induced liver fibrosis was investigated in a mouse model. RESULTS Treatment with APS-1 increased serum levels of interleukin (IL)-12 and interferon (IFN)-γ, increased superoxide dismutase (SOD) activity, and decreased levels of IL-13 and IL-5, and hyaluronidase activity. Moreover, immunohistochemical analysis revealed that the collagen content of hepatic tissue of APS-1-treated mice, including that of collagen I, II, and IV, was dramatically decreased. Furthermore, our data showed that combined treatment of APS-1 with praziquantel had more pronounced effects than treatment with either APS-1 or praziquantel alone. CONCLUSIONS Our findings suggest that the treatment using APS-1 in combination with praziquantel attenuated S. japonicum egg-induced hepatic fibrosis, and possessed potent hepatoprotective activity.

UCA1 functions as a competing endogenous RNA to suppress epithelial ovarian cancer metastasis.

Urothelial cancer associated 1 (UCA1) is an example of functional long noncoding RNAs involved in many biologic processes. However, little is known about the association between UCA1 expression and metastasis in epithelial ovarian cancer (EOC). Findings of this study confirmed that not only UCA1 was aberrantly upregulated in EOC tissues and cells, but also correlated with status of lymph node metastasis and FIGO stage. Furthermore, univariate and multivariate analyses showed that UCA1 was a prognostic factor for overall survival in EOC patients. In vitro, knockdown of UCA1 reduced the invasion and migration ability of EOC cells. The results showed that UCA1 could function as an endogenous sponge by directly binding to miR-485-5p. Depletion of UCA1 was involved in the downregulation of matrix metallopeptidase 14 (MMP14) expression, a target gene of miR-485-5p. In conclusion, our work indicates that UCA1 is a new prognostic biomarker for EOC, establishing a novel connection among UCA1, miR-485-5p, and MMP14 in EOC metastasis.

[Gynecological malignant tumor related multiple primary malignant neoplasms: clinical analysis of 30 cases].

OBJECTIVE: To investigate the clinical features of gynecological malignant tumor related multiple primary malignant neoplasms (MPMN). METHODS: Apply retrospective and comprehensive analysis to the clinical data of 30 patients with gynecological malignant tumor related MPMN. RESULTS: Synchronous MPMN were found in 9 patients. Their average age was 50.2 years old and their median age was 49 years old. The neoplasms were located at ovary, uterus, cervix, breast and intestine. Metachronous MPMN were found in 21 patients. Their average age was 57.7 and their median age was 57 years old. The median interval between the first and the second primary malignant neoplasm was 4.0 years. The neoplasms were located at breast, ovary, uterus, gastrointestinal tract, uterine cervix, lung etc. In 30 cases, 26 of them were treated by surgical operation and further adjunctive treatment of chemotherapy and (or) radiotherapy was conducted as per the neoplasm staging and its pathological results. The rest 4 patients (first primary malignant neoplasms were excised from 3 of them and another one was not treated by surgical operation) received adjunctive treatment of chemotherapy and (or) radiotherapy. Followed ups, which varied from 6 to 60 months, were made to 29 patients and 20 out of the 29 were alive.5-year survival rate of patients with gynecological malignant tumor related MPMN was 47.8%, 2-year survival rate was 73.9%, and 1-year survival rate was 88.6%. CONCLUSION: Pay more attention to the patients with gynecological malignant tumor related MPMN, examine the high-risk patients with malignant tumor comprehensively, identify whether it is recurrence, metastasis or new growth of malignant neoplasm, and further ensure early diagnosis and proper treatment, avoiding misdiagnosis and missed diagnosis.

[Seasonal changes of glycogen level in Oncomelania hupensis].

Oncomelania hupensis snails were collected from Qingyi River of Wuhu City from August 2012 to July 2013. Livers and pedal muscles of snails were dissected. Anthrone colorimetric method was used to evaluate the glycogen concentrations of whole-body, liver and muscle. The concentration of whole-body and liver glycogen decreased from September to next June. The whole body glycogen content in female (0.55 microg/mg) and male (0.88 microg/mg) snails was the lowest in June and April, respectively. The mean whole-body glycogen concentration in females and males was 2.99 and 3.39 microg/mg, respectively. Liver glycogen concentration was lowest in May (female = 0.29 microg/mg, male = 0.22 microg/mg), and reached peak level in August (female = 2.49 microg/mg, male = 2.78 microg/mg). The average liver glycogen concentration in female and male snails was 1.09 and 0.89 microg/mg, respectively. The muscle glycogen concentration gradually decreased from February to June, the lowest was found in June (female = 0.25 microg/mg, male = 0.41 microg/mg), and reached peak level in December (female =16.59 microg/mg, male = 10.06 microg/mg). The average muscle glycogen concentration in female and male snails was 799 and 605 microg/mg, respectively. There was a positive linear correlation between whole-body and liver glycogen concentrations (P < 0.05), and both of them had the similar trend in their monthly change. A positive linear correlation was found among whole-body, liver and muscle glycogen concentrations (P < 0.05).

Comparison of poly (ADP-ribose) polymerase inhibitors (PARPis) as maintenance therapy for newly-diagnosed and platinum-sensitive recurrent ovarian cancer with BRCA mutational status: a systematic review and network meta-analysis.

BACKGROUND: Poly(adenosine diphosphate [ADP]-ribose) polymerase inhibitors (PARPi) treatment for ovarian cancer (OC) are ever-changing. This study aimed to compare the efficacy and overall safety of available PARPi as maintenance therapy for BRCA mutation status in patients with newly diagnosed and platinum-sensitive recurrent (PSR) OC patients. RESEARCH DESIGN AND METHODS: Relevant RCTs were systematically retrieved from PubMed and Embase until 31 May 2022. Progression-free survival (PFS) and overall survival (OS) based on BRCA mutation status and adverse events (AEs) regardless of mutation were efficacy and safety endpoints. RESULTS: In newly diagnosed BRCAm-OC patients, olaparib (HR: 0.33; 95% confidence interval [CI]: 0.25, 0.43) and other PARPis [niraparib (HR: 0.40; 95% CI: 0.29, 0.55), rucaparib (HR: 0.40; 95% CI: 0.21, 0.76) and veliparib (HR: 0.44; 95% CI: 0.28, 0.69)] had a statistically significant effect on PFS versus placebo. In BRCAm-PSROC patients, Olaparib exhibited significant benefit (HR: 0.69; 95% CI: 0.54, 0.88) for OS compared to other PARPis. In BRCAwt-PSR OC patients, Olaparib showed a favorable OS benefit than other PARPis (HR: 0.84; 95% CI: 0.57,1.22). Overall, safety profile of all PARPis was acceptable. CONCLUSION: All PARPis showed significant benefit, with olaparib showing greater benefit in newly diagnosed and PSR OC women. REGISTRATION: CRD42021288932.

Beyond monotherapy: An era ushering in combinations of PARP inhibitors with immune checkpoint inhibitors for solid tumors.

The introduction of PARP inhibitors (PARPis) and immune checkpoint inhibitors (ICIs) has marked a significant shift in the treatment landscape for solid tumors. Emerging preclinical evidence and initial clinical trials have indicated that the synergistic application of PARPis and ICIs may enhance treatment efficacy and potentially improve long-term patient outcomes. Nonetheless, how to identify specific tumor types and molecular subgroups most likely to benefit from this combination remains an area of ongoing research. This review thoroughly examines current studies on the co-administration of PARPis and ICIs across various solid tumors. It explores the underlying mechanisms of action, evaluates clinical efficacy, identifies potential responder populations, and delineates common adverse events alongside strategic management approaches. The aim is to offer a detailed understanding of this combination therapy, potentially guiding future therapeutic strategies for solid tumors.

[Extraction and morphological observation of Oncomelania hupensis haemocytes].

Haemocytes were collected from Oncomelania hupensis snails by using tissue disruption and filtration method, stained by Giemsa and methylene blue, respectively, and observed under microscope. Number of haemocytes from one snail was counted. Out of 54 haemocytes, 3 types of cells were found: big round cells with particles, small round cells with oval nuclei and spindle cells with oblong nuclei. The diameter of big round cells with particles and small round cells with oval nuclei was (21.59-31.97) and (13.24-20.77) microm, respectively. Spindle cells with oblong nuclei was about (17.60-25.47) microm x (27.19-30.25) microm. The nucleocytoplasmic ratio of the above three type cells was 0.38, 0.44 and 0.38, and occupied 35.95% (19/54), 12.42% (28/54) and 51.63% (7/54), respectively. About 194 600 haemocytes were filtered from one single snail. It means that this filtration method is an effective one to extract haemocytes from O. hupensis.

Clinical characteristics and serum CA19-9 combined with HE4 are valuable in diagnosing endometriosis-associated ovarian cancer.

OBJECTIVE: Endometriosis-associated ovarian cancer (EAOC) is difficult to diagnose because of its low incidence, uncertain risk factors, and the absence of effective markers. This study aimed to investigate the clinical characteristics of EAOC and identify useful serological markers. METHODS: We retrospectively studied the clinical characteristics of patients with EAOC and ovarian endometriosis, obtained between January 1, 2011 and October 31, 2021. Univariate and multivariate logistic regression analyses were used to explore the relationship between clinical characteristics and EAOC. Receiver operating characteristic curves were applied to access the diagnostic value of serological markers in EAOC. RESULTS: In total, the clinical characteristics of 220 patients were obtained; 44 with EAOC and 176 with ovarian endometriosis. EAOC patients were older (46.20 vs. 36.26 years, P < 0.001) and had larger tumors (9.10 vs. 6.73 cm, P = 0.003) together with higher CA19-9 (21.44 vs. 4.72 U/mL, P < 0.001) and HE4 levels (62.35 vs. 44.19 pmol/L, P < 0.001) when compared with ovarian endometriosis patients. Multivariate analysis showed that HE4 greater than 59.7 pmol/L, CA19-9 greater than 8.5 U/mL, age 42 years or older, and tumor length 9.2 cm or longer were independent risk factors for EAOC. Significantly, CA19-9 combined with HE4 had high sensitivity (72.73%) and specificity (78.41%) in diagnosing EAOC. CONCLUSION: Age over 42 years, large ovarian tumor, serum CA19-9 and HE4 are valuable in the diagnosis of EAOC.

ALKBH5 activates FAK signaling through m6A demethylation in ITGB1 mRNA and enhances tumor-associated lymphangiogenesis and lymph node metastasis in ovarian cancer.

Background: Lymph node (LN) metastasis is common in patients with epithelial ovarian cancer (EOC) and is associated with poor prognosis. Tumor-associated lymphangiogenesis is the first stage of LN metastasis. Research on lymphangiogenesis and lymph node metastases can help develop new anti-LN-targeted therapies. Aberrant N6-methyladenosine (m6A) modifications have been reported to be linked to LN metastasis in several cancers, however, their role in EOC lymphangiogenesis and LN metastasis remains unclear. Methods: m6A levels in EOC tissues with or without LN metastases were evaluated by dot blot analysis. Real-time polymerase chain reaction (PCR) and immunofluorescence were used to examine the expression of m6A-related enzymes. Additionally, in vitro and in vivo functional studies were performed to discover the importance of the AlkB homolog 5 (ALKBH5) gene in EOC lymphatic metastasis. To identify the downstream target genes regulated by ALKBH5, we performed RNA pulldown, RNA-binding protein immunoprecipitation-quantitative PCR, co-immunoprecipitation, m6A-modified RNA immunoprecipitation-quantitative PCR, and luciferase reporter assays. Results: m6A modification was reduced in ovarian cancers with LN metastases. ALKBH5 overexpression increased tumor-associated lymphangiogenesis and LN metastasis both in vitro and in vivo. ALKBH5 overexpression also reversed the m6A modification in ITGB1 mRNA and suppressed the YTHDF2 protein-mediated m6A-dependent ITGB1 mRNA degradation, which resulted in increased expression of ITGB1 and phosphorylation of the focal adhesion kinase (FAK) and Src proto-oncogene proteins, thereby increasing LN metastasis. Furthermore, hypoxia induced the expression of hypoxia inducible factor 1 subunit alpha, which increased ALKBH5 expression and enhanced LN metastasis in EOC. Conclusions: The ALKBH5/m6A-ITGB1/FAK signalling axis is important in ovarian cancer lymphangiogenesis and LN metastasis. Antibodies that block ITGB1 and FAK kinase-inhibitors are promising anti-metastatic agents.

A Low Dose of Ouabain Alters the Metabolic Profile of Adult Rats Experiencing Intrauterine Growth Restriction in a Sex-Specific Manner.

Intrauterine growth restriction (IUGR) increases the risk of type 2 diabetes mellitus (T2DM) and metabolic diseases. The pancreas of fetuses with IUGR is usually characterized by pancreatic dysplasia and reduced levels of insulin secretion caused by the diminished replication of ß-cells. Previous studies showed that a low dose of ouabain could reduce the apoptosis of embryonic nephric cells during IUGR and partially restore the number of nephrons at birth. The rescued kidneys functioned well and decreased the prevalence of hypertension. Thus, we hypothesized that ouabain could rescue pancreatic development during IUGR and reduce the morbidity of T2DM and metabolic diseases. Maternal malnutrition was used to induce the IUGR model, and then a low dose of ouabain was administered to rats with IUGR during pregnancy. Throughout the experiment, we monitored the pattern of weight increase and evaluated the metabolic parameters in the offspring in different stages. Male, but not female, offspring in the IUGR group presented catch-up growth. Ouabain could benefit the impaired glucose tolerance of male offspring; however, this desirable effect was eliminated by aging. The insulin sensitivity was significantly impaired in male offspring with IUGR, but it was improved by ouabain, even during old age. However, in the female offspring, low birth weight appeared to be a beneficial factor even in old age; administering ouabain exacerbated these favorable effects. Our data suggested that IUGR influenced glucose metabolism in a sex-specific manner and ouabain treatment during pregnancy exerted strongly contrasting effects in male and female rats.

[Extraction of agglutinin from Oncomelania hupensis and its haemagglutination activity].

OBJECTIVE: To explore the extraction methods of agglutinin from Oncomelania hupensis snail and study its haemagglutination activity. METHODS: Protein obtained by ammonium sulfate fractionation precipitation with 20%-100% saturation of ammonium sulfate. Its haemagglutination activity was determined by rabbit erythrocytes. The precipitation which could agglutinate rabbit erythrocytes was diluted with 2.5 mg/ml D-galactose, D-fructose, D-glucose, saccharose, maltose and lactose, respectively, and then their haemagglutination activity was tested. Snail agglutinin were incubated at different temperatures (25-90 degrees C) and assayed for agglutinating activity. The effect of pH on the haemagglutination activity was determined by using the PBS buffer at different pH values (3.0-10.0). RESULTS: Oncomelania snail agglutinin exhibited high haemagglutination activity in 20%-40% saturated ammonium sulfate pellet. Lactose and galactose could inhibit the haemagglutination activity of snail agglutinin. The agglutinin showed maximum activity at pH 7.0. In temperature range of 30-70 degrees C, the haemagglutination activity decreased with increasing temperature, and all activity lost beyond 80 degrees C. CONCLUSION: Galactose/lactose specific agglutinin exists in Oncomelania snail, its haemagglutination activity is affected by pH and temperature.

[Isolation and purification of Oncomelania hupensis agglutinin and determination of its molecular weight].

OBJECTIVE: To isolate and purify agglutinin from Oncomelania hupensis snail and determine its molecular weight. METHODS: Agglutinin was preliminarily isolated from snail tissue homogenate by 0%-40% saturated ammonium sulfate, and then successively purified with Sephadex G-75 gel filtration and Sepharose 4B affinity chromatography. Bradford assay was used to determine the protein content. The agglutination activity was determined by rabbit erythrocytes. The purity of agglutinin preparations was assessed by SDS-PAGE. The molecular weight of agglutinin subunit was determined by Sephadex G-75 gel filtration. RESULTS: The specific activity of snail tissue homogenate was 21.74 titer/mg. After ammonium sulfate precipitation, Sephadex G-75 gel filtration and Sepharose 4B affinity chromatography, the specific activity of snail agglutinin from the homogenate solution increased to 61.93 titer/mg, 75.89 titer/mg and 963.86 titer/mg, respectively. SDS-PAGE analysis indicated that snail agglutinin (M, 53,000) was purified by Sephadex G-75 gel filtration and Sepharose 4B chromatography. The molecular weight of the snail agglutinin produced by Sephadex G-75 gel filtration was Mr 78,000. CONCLUSION: Combined use of salt fractionation, gel filtration and affinity chromatography can be efficient for extraction and purification of agglutinin from Oncomelania hupensis species. The snail agglutinin is characterized as mono subunit protein with a molecular weight of Mr 78,000.