ZFP57 promotes ovarian cancer progression by transcriptionally regulating BRCA1 and managing G1 checkpoint.

Ovarian cancer (OC) which is one of the frequently-occurring gynecologic malignant tumors, endangers the health of women. The zinc finger protein 57 (ZFP57) plays crucial functions during the progression of cancer and is reported as a prognostic and therapeutic candidate in a variety of cancer. However, the biological function as well as the underlying mechanism of ZFP57 during OC progression remains unknown. Here, ZFP57 expression was found prominently increased in OC tissues and correlated with the prognosis of OC patients. Knock down of ZFP57 in OC cells inhibited the cell proliferation and migration, and also arrested the cells at G1 phase as well as accelerated the apoptosis. Additionally, ZFP57 transcriptionally regulated BRCA1 expression in OC, indicating that ZFP57 may affect BRCA1 mediated G1 checkpoint to regulate the cell cycle of OC cells and further influence the progression of OC. Taken together, our present study discovered a novel function of ZFP57 in OC, suggesting that ZFP57 could be potentially treated as a prognostic biomarker and therapeutic target for OC patients.

Effects of two blood purification treatments combined with immunosuppressants on GM-CSF and CXCL16 levels in patients with lupus nephritis.

OBJECTIVES: This study aimed to investigate the therapeutic effects of two different blood purification treatments combined with immunosuppressants on patients with lupus nephritis (LN) and their effects affecting granulocyte-macrophage colony-stimulating factor (GM-CSF) and CXC chemokine ligand-16 (CXCL16) levels. METHODS: Ninety patients with LN admitted to our hospital were enrolled and randomly assigned into groups A and B. Group A was treated with continuous veno-venous hemofiltration (CVVH) combined with conventional medical treatment (CMT, n = 40, including 23 females and 17 males), whereas group B was treated with intermittent hemodialysis combined with conventional medical treatment (n = 50, including 35 females and 15 males). Both groups received prednisone and cyclophosphamide. RESULTS: GM-CSF and CXCL16 levels in the two groups were significantly reduced after treatment (P < 0.05); and GM-CSF level in group A was significantly lower than that in group B (P < 0.05, -1.261 to -0.8395), and CXCL16 level in group A was significantly lower than that in group B (P < 0.05, -0.5745 to -0.4355). There was no significant difference in general data between the two groups (P > 0.05). After treatment, the SLEDAI scores were significantly decreased in both groups, and were significantly lower in group A than in group B (P < 0.05, -1.816 to -0.1241). CONCLUSION: CVVH combined with conventional medical treatment is more effective than intermittent hemodialysis combined with conventional medical treatment, and is easier to remove GM-CSF and CXCL16.

Genomic characterization of Chinese ovarian clear cell carcinoma identifies driver genes by whole exome sequencing.

Little is known about the genetic alterations characteristic of ovarian clear cell carcinoma (OCCC). Our aim was to identify targetable genomic alterations in this type of cancer. Forty-two OCCC formalin-fixed, paraffin-embedded (FFPE) tissue samples were analyzed by whole-exome sequencing (WES), and 74 FFPE tissue samples underwent targeted sequencing (TS) to confirm the relevant driver mutations. Cell proliferation was assessed by cell counting kit-8 (CCK8) assays. In the 42 samples, ARID1A (64.3%) and PIK3CA (28.5%) were frequently mutated, as were PPP2R1A (11.9%), PTEN (7.1%) and KRAS (4.8%), which have been reported in previous OCCC studies. We also detected mutations in MUC4 (28.6%), MAGEE1 (19%), and ARID3A (16.7%); associations with these genes have not been previously reported. The functional protein-activated pathways were associated with proliferation and survival (including the PI3K/AKT, TP53, and ERBB2 pathways) in 83% of OCCCs and with chromatin remodeling in 71% of OCCCs. Patients with alterations in MAGEE1 (64% in the targeted sequencing cohort) had worse clinical outcomes (log-rank p < 0.05). A functional study revealed that two MAGEE1 mutants, one lacking two MAGE domains and the other containing two MAGE domains, significantly decreased the proliferative capacity of OCCC cells. We successfully identified novel genetic alterations in OCCC using whole-exome sequencing and targeted sequencing of OCCC patient samples and potential therapeutic targets for the treatment of this malignancy.

High expression of ZNF93 promotes proliferation and migration of ovarian cancer cells and relates to poor prognosis.

Ovarian cancer (OC) is most common type of gynecologic cancer and is frequently lethal. It is important to determine the pathologic mechanisms underlying OC. ZNF93 is a member of the zinc finger protein family. Abnormal expression of ZNF93 has been observed in various tumor cells. However, its clinical significance and biologic function in ovarian cancer remain unclear. In the present study, we established that ZNF93 expression was highly up-regulated in OC samples and was closely correlated with clinical stage, indicating poor prognosis. We then established that ZNF93 promoted OC cell proliferation and migration. The results of our study may provide insight into the use of ZNF93 as a marker of clinical outcome and as a potential therapeutic target in OC.

Structure determination of CAMP factor of Mobiluncus curtisii and insights into structural dynamics.

Bacterial vaginosis (BV) is a common type of vaginal inflammation caused by a proliferation of pathogenic bacteria, among which Mobiluncus curtisii. In our previous studies on M. curtisii genome, we identified the presence of a genomic fragment encoding a 25 kDa pore-forming toxin, the CAMP factor, which is known to be involved in the synergistic lysis of erythrocytes namely CAMP reaction. However, whether this hypothetical gene product has hemolytic activity is unknown. Moreover, its relative structure and function are not yet solved. Here we found that the M. curtisii CAMP factor is a monomer at pH 4.4 and oligomer at pH > 4.6. Hemolysis assays showed that M. curtisii CAMP factor could lyse sheep red blood cells efficiently in pH 5.4-7.4. Negative staining electron microscope analysis of the CAMP factor revealed ring-like structures at pH above 4.6. Additionally, the crystal structure of M. curtisii CAMP factor, determineded at 1.85 Å resolution, reveals a 5 + 3 helix motif. Further functional analysis suggested that the structural rearrangement of the N-terminal domain might be required for protein function. In conclusion, this structure-function relationship study of CAMP factor provides a new perspective of the M. curtisii role in BV development.

Facile Synthesis of a H2O2-Responsive Alternating Copolymer Bearing Thioether Side Groups for Drug Delivery and Controlled Release.

A novel amphiphilic alternating copolymer with thioether side groups (P(MSPA-a-EG)) was synthesized through an amine-epoxy click reaction of 3-(methylthio)propylamine (MSPA) and ethylene glycol diglycidyl ether. P(MSPA-a-EG) was characterized in detail by nuclear magnetic resonance (NMR), gel permeation chromatography, Fourier transformed infrared, differential scanning calorimeter, and thermogravimetric analysis to confirm the successful synthesis. Due to its amphiphilic structure, P(MSPA-a-EG) could self-assemble into spherical micelles with an average diameter of about 151 nm. As triggered by H2O2, theses micelles could disassemble because hydrophobic thioether groups are transformed to hydrophilic sulfoxide groups in MSPA units. The oxidant disassemble process of micelles was systemically studied by dynamic light scattering, transmission electron microscopy, and 1H NMR measurements. The MTT assay against NIH/3T3 cells indicated that P(MSPA-a-EG) micelles exhibited good biocompatibility. Furthermore, they could be used as smart drug carriers to encapsulate hydrophobic anticancer drug doxorubicin (DOX) with 4.90% drug loading content and 9.81% drug loading efficiency. In vitro evaluation results indicated that the loaded DOX could be released rapidly, triggered by H2O2. Therefore, such a novel alternating copolymer was expected to be promising candidates for controlled drug delivery and release.

Structure determination of the CAMP factor of Streptococcus agalactiae with the aid of an MBP tag and insights into membrane-surface attachment.

CAMP factor is a unique α-helical bacterial toxin that is known for its co-hemolytic activity in combination with staphylococcal sphingomyelinase. It was first discovered in the human pathogen Streptococcus agalactiae (also known as group B streptococcus), but homologous genes have been found in many other Gram-positive pathogens. In this study, the efforts that led to the determination of the first structure of a CAMP-family toxin are reported. Initially, it was possible to produce crystals of the native protein which diffracted to near 2.45 Šresolution. However, a series of technical obstacles were encountered on the way to structure determination. Over a period of more than five years, many methods, including selenomethionine labeling, mutations, crystallization chaperones and heavy-atom soaking, were attempted, but these attempts resulted in limited progress. The structure was finally solved using a combination of iodine soaking and molecular replacement using the crystallization chaperone maltose-binding protein (MBP) as a search model. Analysis of native and MBP-tagged CAMP-factor structures identified a conserved interaction interface in the C-terminal domain (CTD). The positively charged surface may be critical for binding to acidic ligands. Furthermore, mutations on the interaction interface at the CTD completely abolished its co-hemolytic activities. This study provides novel insights into the mechanism of the membrane-permeabilizing activity of CAMP factor.

Crystal structure of the Streptococcus agalactiae CAMP factor provides insights into its membrane-permeabilizing activity.

Streptococcus agalactiae is an important human opportunistic pathogen that can cause serious health problems, particularly among newborns and older individuals. S. agalactiae contains the CAMP factor, a pore-forming toxin first identified in this bacterium. The CAMP reaction is based on the co-hemolytic activity of the CAMP factor and is commonly used to identify S. agalactiae in the clinic. Closely related proteins are present also in other Gram-positive pathogens. Although the CAMP toxin was discovered more than a half century ago, no structure from this toxin family has been reported, and the mechanism of action of this toxin remains unclear. Here, we report the first structure of this toxin family, revealing a structural fold composed of 5 + 3-helix bundles. Further analysis by protein truncation and site-directed mutagenesis indicated that the N-terminal 5-helix bundle is responsible for membrane permeabilization, whereas the C-terminal 3-helix bundle is likely responsible for host receptor binding. Interestingly, the C-terminal domain inhibited the activity of both full-length toxin and its N-terminal domain. Moreover, we observed that the linker region is highly conserved and has a conserved DLXXXDXAT sequence motif. Structurally, this linker region extensively interacted with both terminal CAMP factor domains, and mutagenesis disclosed that the conserved sequence motif is required for CAMP factor's co-hemolytic activity. In conclusion, our results reveal a unique structure of this bacterial toxin and help clarify the molecular mechanism of its co-hemolytic activity.

CTHRC1 as a novel biomarker in the diagnosis of cervical squamous cell carcinoma.

To verify whether the collagen triple helix repeat containing 1 (CTHRC1) can be used as a potential biomarker in diagnosis of cervical cancer, by evaluating the expression level of CTHRC1 in cervical squamous cell carcinoma patients. In this study, CTHRC1 expression in cervical squamous cell carcinoma, CIN (cervical intraepithelial neoplasia) and healthy cervical squamous epithelium were measured by immunohistochemistry. The serum levels of CTHRC1 and SCC-Ag within the all three groups were performed using ELISA. In addition, the ROC curve of CTHRC1 and SCC-Ag as well as combined CTHRC1 and SCC-Ag was demonstrated and analyzed. CTHRC1 was significantly overexpressed in cervical squamous cell carcinoma compared with CIN and healthy control group. And CTHRC1 concentration in serum of cervical squamous cell carcinoma group was also remarkably higher than that in other two groups. The ROC curve showed AUC of CTHRC1 and SCC-Ag was 0.665±0.034 and 0.878±0.027, the sensitivity of them were 57.1% and 77.6%, and the specificity of them were 85.4% and 86%, respectively. Furthermore, AUC of combined CTHRC1 and SCC-Ag was 0.879±0.027, sensitivity was 87.2% and specificity were 84%. Our study indicated that CTHRC1 was highly upregulated not only in the tissue but also in the serum of cervical squamous cell carcinoma patients, which pointed out it can be used as a novel prognostic and metastatic biomarker of cervical squamous cell carcinoma. And combined SCC-Ag and CTHRC1 serological detection may have potential value in the early diagnosis of cervical squamous cell carcinoma and CIN.