Chitosan-coated miconazole as an effective anti-inflammatory agent for the treatment of postoperative infections in obstetrics and vaginal yeast infection control on in vitro evaluations.

People with immune deficiency are at risk of developing infections caused by several bacterial and fungal species. In this work, chitosan-coated miconazole was developed by a simple sol-gel method. Miconazole is considered an effective drug to treat vaginal infection-causing bacteria and fungi. The coating of chitosan with miconazole nitrate showed the highest drug loading efficiency (62.43%) and mean particle size (2 µm). FTIR spectroscopic analysis confirmed the entrapment of miconazole nitrate into chitosan polymer. The antifungal result demonstrated that MN@CS microgel possessed notable anti-Aspergillus fumigatus and Candida albicans activity in lower doses. Antibacterial activity results revealed excellent bacterial growth inhibition of MN@CS microgel towards human skin infectious pathogens Escherichia coli and Staphylococcus aureus. The biocompatibility studies of In vitro cell viability and Artemia salina lethality assay suggested that MN@CS microgel is more biosafe and suitable for human external applications. In the future, it will be an efficient anti-inflammatory agent for the treatment of vaginal infections.

Genome-Wide Identification, Characterization, and Expression Analysis of Tubby-like Protein (TLP) Gene Family Members in Woodland Strawberry (Fragaria vesca).

Tubby-like proteins (TLPs) play important roles in plant growth and development and in responses to abiotic stress. However, TLPs in strawberry remain poorly studied. In this study, eight TLPs were identified in woodland strawberry (Fragaria vesca subspecies vesca 'Ruegen'). Protein structure analysis revealed that the structure of FvTLPs is highly conserved, but evolutionary and gene structure analyses revealed that the evolutionary pattern of FvTLP family members differs from that of their orthologous genes in Arabidopsis, poplar, and apple. Subcellular localization assays revealed that FvTLPs were localized to the nucleus and plasma membrane. FvTLPs showed no transcriptional activity. Yeast two-hybrid assays revealed that FvTLPs interact with specific FvSKP1s. The expression patterns of FvTLPs in different tissues and under various abiotic stresses (salt, drought, cold, and heat) and hormone treatments (ABA (abscisic acid) and MeJA (methyl jasmonate)) were determined. The expression patterns of FvTLPs indicated that they play a role in regulating growth and development and responses to abiotic stress in F. vesca. The GUS (beta-glucuronidase) activity of FvTLP1pro::GUS plants in GUS activity assays increased under salt and drought stress and abscisic acid treatment. The results of this study provide new insights into the molecular mechanisms underlying the functions of TLPs.

Combined transcriptomic and metabolomic analysis reveals a role for adenosine triphosphate-binding cassette transporters and cell wall remodeling in response to salt stress in strawberry.

Strawberry (Fragaria × ananassa Duch) are sensitive to salt stress, and breeding salt-tolerant strawberry cultivars is the primary method to develop resistance to increased soil salinization. However, the underlying molecular mechanisms mediating the response of strawberry to salinity stress remain largely unknown. This study evaluated the salinity tolerance of 24 strawberry varieties, and transcriptomic and metabolomic analysis were performed of 'Sweet Charlie' (salt-tolerant) and 'Benihoppe' (salt-sensitive) to explore salt tolerance mechanisms in strawberry. Compared with the control, we identified 3412 differentially expressed genes (DEGs) and 209 differentially accumulated metabolites (DAMs) in 'Benihoppe,' and 5102 DEGs and 230 DAMs in 'Sweet Charlie.' DEGs Gene Ontology (GO) enrichment analyses indicated that the DEGs in 'Benihoppe' were enriched for ion homeostasis related terms, while in 'Sweet Charlie,' terms related to cell wall remodeling were over-represented. DEGs related to ion homeostasis and cell wall remodeling exhibited differential expression patterns in 'Benihoppe' and 'Sweet Charlie.' In 'Benihoppe,' 21 ion homeostasis-related DEGs and 32 cell wall remodeling-related DEGs were upregulated, while 23 ion homeostasis-related DEGs and 138 cell wall remodeling-related DEGs were downregulated. In 'Sweet Charlie,' 72 ion homeostasis-related DEGs and 275 cell wall remodeling-related DEGs were upregulated, while 11 ion homeostasis-related DEGs and 20 cell wall remodeling-related DEGs were downregulated. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed only four KEGG enriched pathways were shared between 'Benihoppe' and 'Sweet Charlie,' including flavonoid biosynthesis, phenylalanine metabolism, phenylpropanoid biosynthesis and ubiquinone, and other terpenoid-quinone biosynthesis. Integrating the results of transcriptomic and metabolomics analyses showed that adenosine triphosphate-binding cassette (ABC) transporters and flavonoid pathway genes might play important roles in the salt stress response in strawberry, and DAMs and DEGs related to ABC transporter and flavonoid pathways were differentially expressed or accumulated. The results of this study reveal that cell wall remodeling and ABC transporters contribute to the response to salt stress in strawberry, and that related genes showed differential expression patterns in varieties with different salt tolerances. These findings provide new insights into the underlying molecular mechanism of strawberry response to salt stress and suggest potential targets for the breeding of salt-tolerant strawberry varieties.

The complete mitochondrial genome sequence of Fragaria orientalis (Rosaceae).

Fragaria orientalis Lozinsk. is valuable germplasm material for cross breeding in Fragaria. In this study, we assembled the complete mitochondrial genome of F. orientalis using a combination of Illumina data and Nanopore data. The mitochondrial genome was 275,143 bp in length, including 29 protein-coding genes, 20 tRNA genes, and three rRNA genes, with a total GC content 45.23%. Seven protein-coding genes contained introns, and three were trans-spliced. Phylogenetic analysis indicated that F. orientalis is making a sister clade to the Amygdaloideae species. The complete mitochondrial genome of F. orientalis reported in this study will improve our understanding of Fragaria evolution.

Correlation Between High Expression of FOXA2 and Improved Overall Survival in Ovarian Cancer Patients.

BACKGROUND The aim of the present work was to evaluate FOXA2 expression in ovarian cancer and to use integrated bioinformatics analysis to correlate it with patient prognosis. MATERIAL AND METHODS FOXA2 expression was evaluated in multiple cancers in The Cancer Genome Atlas database. A protein-protein interaction (PPI) network relevant to FOXA2 was constructed using the Search Tool for Retrieval of Interacting Genes/Proteins (STRIN). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed of FOXA2 and relevant genes. Correlations between overall survival (OS), disease-free survival, and FOXA2 expression were evaluated. An immunohistochemical assay (IHC) was used to test for FOXA2 protein expression in 79 ovarian cancer specimens. RESULTS FOXA2 mRNA was upregulated in colorectal, stomach, liver, and endometrial cancers. In the PPI network, 21 protein nodes and 533 edges were constructed with a local clustering coefficient of 0.698, which indicated significant PPI enrichment (P<0.01). FOXA2 and relevant genes were mainly enriched in the signaling pathways regulating pluripotency of stem cells, cancer, and AMPK. A survival analysis indicated that OS was significantly longer in patients with higher versus lower FOXA2 protein expression (HR=0.73, P<0.01). The IHC assay showed that the FOXA2 protein was mainly positively expressed in the nucleoplasm of tumor cells with brown-yellow staining. Of the 79 ovarian cancer samples, 31 (39.2%) highly expressed FOXA2. The FOXA2 gene was correlated with International Federation of Gynecology and Obstetrics staging and with lymph node metastasis (both P<0.05). CONCLUSIONS Upregulation of the FOXA2 gene was correlated with improved OS in patients with ovarian cancer and it can be used as a prognostic biomarker and potential treatment target.

Drift Reduction of a 4-DOF Measurement System Caused by Unstable Air Refractive Index.

Laser beam drift greatly influences the accuracy of a four degrees of freedom (4-DOF) measurement system during the detection of machine tool errors, especially for long-distance measurement. A novel method was proposed using bellows to serve as a laser beam shield and air pumps to stabilize the refractive index of air. The inner diameter of the bellows and the control mode of the pumps were optimized through theoretical analysis and simulation. An experimental setup was established to verify the feasibility of the method under the temperature interference condition. The results indicated that the position stability of the laser beam spot can be improved by more than 79% under the action of pumping and inflating. The proposed scheme provides a cost-effective method to reduce the laser beam drift, which can be applied to improve the detection accuracy of a 4-DOF measurement system.

Modeling and Optimal Design for a High Stability 2D Optoelectronic Angle Sensor.

The structural deformations caused by environmental changes in temperature, vibration, and other factors are harmful to the stability of high precision measurement equipment. The stability and optimal design method of a 2D optoelectronic angle sensor have been investigated in this study. The drift caused by structural deformations of the angle sensor has been studied and a drift error model has been achieved. Key components sensitive to thermal and vibrational effects were identified by error sensitivity analysis and simulation. The mounts of key components were analyzed using finite element analysis software and optimized based on the concept of symmetric structures. Stability experiments for the original and optimized angle sensors have been carried out for contrast. As a result, the stability of the optimized angle sensor has been improved by more than 63%. It is verified that the modeling and optimal design method is effective and low-cost, which can also be applied to improve the stability of other sensors with much more complex principles and structures.

Axillary lymph node metastasis detection by magnetic resonance imaging in patients with breast cancer: A meta-analysis.

BACKGROUND: The study was conducted to evaluate the diagnostic performance of magnetic resonance imaging (MRI) for the detection of axillary lymph node metastasis in patients with breast cancer. METHODS: PubMed, Medline, Web of Science, Cochrane Embase, Chinese Biomedical Literature, and China National Knowledge Infrastructure databases were searched for open published studies relevant to the use of MRI for the detection of axillary lymph node metastasis in breast cancer patients. The pooled diagnostic sensitivity, specificity, and the symmetric receiver operating characteristic (SROC) curve was calculated by combining the individual data extracted from 26 included studies. RESULTS: The pooled diagnostic sensitivity and specificity of MRI to detect axillary lymph node metastasis in patients with breast cancer were 0.77 (95% confidence interval [CI] 0.75-0.80) and 0.90 (95% CI 0.89-0.91), respectively. The pooled positive and negative likelihood ratios were 7.67 (95% CI 5.09-11.53) and 0.23 (95% CI 0.17-0.32), respectively, by random effect method. The area under the SROC curve was 0.93 for MRI to detect axillary lymph node metastasis in breast cancer patients. CONCLUSION: With high sensitivity, specificity, and area under the curve, MRI is an effective method to differentiate metastatic axillary lymph node in breast cancer patients, which can provide useful information for surgical procedure selection.

Knockdown of SLC39A7 suppresses cell proliferation, migration and invasion in cervical cancer.

Cervical cancer is the fourth leading cause of malignancy related mortality in women worldwide. SLC39A7 (ZIP7) is a zinc transporter that plays a key role in intestinal epithelial self-renewal. However, whether or not SLC39A7 is involved in human cervical cancer remains unclear. In this study, we investigated the effects of SLC39A7 in cervical cancer in vitro and elucidate related underlying mechanisms. Using Oncomine data analysis, we first found SLC39A7 is commonly upregulated in cervical cancer tissues in comparison with corresponding normal controls. The in vitro experiments indicated that silencing of SLC39A7 expression resulted in decreased cell proliferation, increased cell apoptosis, and attenuated migratory and invasive ability using CCK-8, colony formation, flow cytometry, transwell assays, respectively in cervical cancer cell lines, HeLa and ME-180 cells. In molecular levels, Western blot further demonstrated that silencing of SLC39A7 significantly upregulated the expression of Bax and E-cadherin, downregulated the expression of Bcl-2 and MMP-2 in both HeLa and ME-180 cells. These findings provide evidence that SLC39A7 plays a positive role in the progression of cervical cancer and its knockdown might be as a potential therapeutic target for cervical cancer treatment.

Siva 1 inhibits proliferation, migration and invasion by phosphorylating Stathmin in ovarian cancer cells.

Ovarian cancer is one of the most common types of gynecologic malignant tumor, with high incidence and high mortality rates. It is difficult to diagnose ovarian cancer early due to the complex structure and function of the ovaries. Siva 1 is a well-known pro-apoptosis protein that functions in multiple types of cancer cells: There are several studies demonstrating that Siva 1 arrests apoptosis and facilitates cancer development in osteosarcoma and non-small cell lung cancer. Whether Siva 1 functions in ovarian cancer remains unknown. In the present study, it was established that Siva 1 was stably overexpressed in ovarian cancer cell lines, and demonstrated that the overexpression of Siva 1 inhibited proliferation, promoted apoptosis and suppressed migration and invasion by facilitating phosphorylation of Stathmin and polymerization of α-tubulin in ovarian cancer cells. These data provide specific novel insights into the molecular mechanism of ovarian cancer, and may be of significance for the clinical diagnosis and therapy of ovarian cancer.

Genome-wide analysis of aberrant gene expression and methylation profiles reveals susceptibility genes and underlying mechanism of cervical cancer.

BACKGROUND: This study aimed to explore the molecular mechanism of cervical cancer (CC) by integrated bioinformatic analyses of gene expression and methylation profiles. METHODS: The gene expression and methylation microarrays in CC samples and normal controls were respectively downloaded from the GEO database. After screening the differentially expressed genes (DEGs) with Limma package and the CC-related methylation sites with CpGassoc package in R language, DEGs with CC-related methylation sites were identified from the intersection of the above two groups of results with 50kb upstream and downstream of a gene as the gene region. Then GO enrichment was performed by GenCLIP2.0 software. Sequentially, analysis of metabolic sub-pathways with pathogenic risk was predicted by iSubpathwayMiner package in R language. RESULTS: A total of 1357 DEGs including 721 up-regulated and 636 down-regulated, as well as 666 CC-related methylation sites were screened out. After being analyzed, 26 DEGs with 35 CC-related methylation sites were identified. EDN3 and EDNRB were significantly involved in a function cluster in GO terms of vein smooth muscle contraction, vascular smooth muscle contraction and phasic smooth muscle contraction. LHX2 and PAX6 were significantly involved in a function cluster in GO terms of telencephalon regionalization and forebrain regionalization. ACOX3, CYP39A1 and DPYS were significantly enriched in 25 sub-pathways of 6 major pathways. CONCLUSIONS: EDN3 and EDNRB might play important roles in the molecular mechanism of CC, and LHX2, ACOX3, CYP39A1 and DPYS might be susceptibility genes and potential risk markers in CC.

KILLER CELL IMMUNOGLOBULIN-LIKE RECEPTOR GENES AND THEIR HLA-C LIGANDS IN HASHIMOTO THYROIDITIS IN A CHINESE POPULATION.

OBJECTIVE: Natural killer (NK) cells serve as primary immune surveillance and are partially regulated by combinations of killer immunoglobulin-like receptors (KIR) and their human leukocyte antigen-C (HLA-C) ligands. Alterations in NK cell activity have been associated with Hashimoto thyroiditis (HT). The aim of this study was to determine whether certain KIR/HLA-C genotype combinations play a role in HT pathogenesis. METHODS: The present study enrolled 107 unrelated HT patients and 108 random healthy individuals in a case-control study. Blood was collected for DNA extraction; typing of KIR genes and HLA-C alleles was performed by polymerase chain reaction with sequence specific primers (PCR-SSP), followed by electrophoresis on agarose gels. RESULTS: Among a panel of KIR2D/HLA-C genotype combinations, the frequency of KIR2DS2/HLA-C1 was significantly increased in HT patients compared to controls (33.64% vs. 12.96%, P<.001). To further analyze the precise genotype, we investigated inhibitory or activating KIR/HLA-C gene pairs when their corresponding activating or inhibitory KIR genes were absent in the 2 groups. Only the frequency of KIR2DS2(-)2DL2/3(+)HLA-C1(+) was significantly decreased in HT patients compared to controls (48.60% vs. 70.37%, P = .001). CONCLUSION: Our data suggest that KIR2DS2/HLA-C1 may correlate with HT pathogenesis. On the contrary, the predominance of KIR2DL2/3/HLA-C1 in the absence of KIR2DS2 suggests a potential inhibitory role in HT pathogenesis. In conclusion, our findings may further elucidate the mechanisms underlying the pathogenesis of HT and other autoimmune diseases. ABBREVIATIONS: HLA-C = human leukocyte antigen-C HT = Hashimoto thyroiditis KIR = killer immunoglobulin-like receptor NK = natural killer PCR = polymerase chain reaction.

Expression of miRNA-1233 in placenta from patients with hypertensive disorder complicating pregnancy and its role in disease pathogenesis.

OBJECTIVE: This study is to investigate the expression of miRNA-1233 in placental tissue from patients with hypertensive disorder complicating pregnancy (HDCP) and its role in disease pathogenesis. METHODS: The expression levels of miRNA-1233 and HoxB3 in placental tissue from HDCP patients and normal control subjects, as well as in the in vitro trophoblast cells, were detected with real-time PCR and Western blot analysis. The proliferation and invasion abilities of trophoblast cells were assessed by the cell counting kit (CCK)-8 and transwell chamber assays, respectively. Dual-luciferase reporter assay was performed to evaluate the interaction between miRNA-1233 and Hoxb3. RESULTS: Real-time PCR showed that, compared with the control group, the expression levels of miRNA-1233 were significantly elevated in placental tissue from HDCP patients. On the other hand, both the mRNA and protein expression levels of HoxB3 were significantly decreased in the HDCP group. Moreover, the mRNA and protein expression levels of HoxB3 were significantly declined by the transfection of miRNA-1233 mimics in trophoblast cells. Bioinformatics analysis and the dual-luciferase reporter gene assay showed that, miRNA-1233 targeted HoxB3 in the 3'-UTR and suppressed the gene expression. In addition, the results from the CCK-8 and transwell chamber assays showed that, the transfection of miRNA-1233 significantly decreased the proliferation and invasion abilities of the trophoblast cells. CONCLUSION: In placental tissue from HDCP patients, up-regulated miR-1233 could suppress the expression of HoxB3, and then inhibit the invasion of trophoblast cells, which might contribute to the disease pathogenesis.

Paclitaxel-resistant HeLa cells have up-regulated levels of reactive oxygen species and increased expression of taxol resistance gene 1.

This study is to establish a paclitaxel (PTX)-resistant human cervical carcinoma HeLa cell line (HeLa/PTX) and to investigate its redox characteristics and the expression of taxol resistance gene 1 (Txr1). HeLa cells were treated with PTX and effects of PTX on cell proliferation were detected through cell counting and the MTT assay. Levels of cellular reactive oxygen species (ROS), reduced glutathione (GSH), and oxidized glutathione (GSSG) as well as the ratio of GSH to GSSG were measured by the 2,7-difluorescein diacetate (DCFH-DA) method and the 5,5'dithiobis(2-nitrobenzoic acid) (DTNB) method. Activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were determined by the nitrite formation method, the molybdate colorimetric method, and the DTNB colorimetric method, respectively. The level of Txr1 mRNA was determined by real-time PCR. Compared with the regular HeLa cells, HeLa/PTX cells were larger in size and had more cytoplasmic granules. The population doubling time for HeLa/PTX cells was 1.32 times of that of HeLa cells (P<0.01). HeLa/PTX cells showed stronger resistance to PTX than HeLa cells with a resistance index of 122.69. HeLa/PTX cells had higher levels of ROS (P<0.01) and Txr1 mRNA (P<0.01), lower level of GSH (P < 0.05), and lower activities of SOD (P<0.01) and GPx (P < 0.05) than HeLa cells. HeLa/PTX cells, with higher levels of ROS and Txr1 mRNA expression, are more resistant to PTX than HeLa cells.

Involvement of mitochondria-caspase pathway in Hemoporfin-mediated cell death.

Hemoporfin is a novel second-generation porphyrin-related photosensitizer for ovarian cancer photodynamic treatment (PDT). The purpose of this study was to investigate the molecular mechanisms of Hemoporfin-mediated photocytotoxicity. Human epithelial ovarian cancer cell line 3AO was incubated with different concentrations of Hemoporfin, and phototoxic effects of Hemoporfin on cells were determined using a Cell Viability Analyzer. Apoptosis or necrosis was determined by flow cytometry analysis using the Annexin V-FITC apoptosis kit. Cellular caspase activation was determined using the fluorescent assay kit for caspase-3 and caspase-9. Rhodamine123 was used as a mitochondrial probe and Lucifer Yellow as a lysosomal probe to investigate the intracellular localization of Hemoporfin in 3AO cancer cells. We demonstrated that both high-dose (30 microg mL(-1)) and low-dose (3 microg mL(-1)) Hemoporfin significantly reduced the viability of ovarian cancer cell 3AO with light illumination, and the photocytotoxicity was dose-dependent (P < 0.01). Using a mitochondrial fluorescence probe, we demonstrated a distinct mitochondrial aggregation in 3AO cells with a low concentration of Hemoporfin. Loss of mitochondrial membrane potential was detected as early as 1 h after Hemoporfin-mediated PDT. PDT with low-dose Hemoporfin predominantly induced apoptosis but not necrosis, and both caspase-3 and caspase-9 were activated. Based on our results, mitochondria play an important role in the Hemoporfin-induced apoptosis, and mitochondria membrane potential loss initiated apoptosis via the activation of caspases. Understanding the mechanisms involved in PDT-mediated apoptosis may improve its therapeutic efficacy and facilitate its transition into the clinic.

Intraperitoneal photodynamic therapy for an ovarian cancer ascite model in Fischer 344 rat using hematoporphyrin monomethyl ether.

With limited treatment options, intraperitoneal spread of ovarian cancer is a common problem leading to high morbidity. Intraperitoneal photodynamic therapy combined with debulking surgery to treat residual disease is an alternative choice for clinicians. Hematoporphyrin monomethyl ether (HMME) is a promising second-generation photosensitizer developed in China. Our study was designed to investigate the phototoxicity of HMME on ovarian cancer. NuTu-19, a cell line derived from adenocarcinoma of Fischer 344 rat, and its allogeneic graft ascites tumor model was used in this study. HMME was confirmed to be localized in cytolysosome, and HMME-based photosensitization induced direct necrosis as well as mitochondria damage. The photocytotoxicity of HMME was both light- and drug dose-dependent and no significant dark cytotoxicity was observed in NuTu-19 cells. With the ascite tumor-bearing Fischer 344 rat model, HMME-based intraperitoneal photodynamic therapy was proved to be useful in improving the prognosis of ovarian cancer. Thus, this study provides evidence that HMME-based photodynamic therapy is an effective adjuvant therapy for ovarian cancer.

Overexpression of CHP2 enhances tumor cell growth, invasion and metastasis in ovarian cancer.

BACKGROUND: Calcineurin B homologous protein isoform 2 (CHP2) was identified to be expressed in various malignant cell lines including ovarian cancer, but not in the normal tissue counterpart. The biological function of CHP2 related to cancer progression is still unknown. MATERIALS AND METHODS: A CHP2-negative human epithelial ovarian cancer cell line OVCAR3 was used for this study. CHP2 was analyzed before and after gene transfection. Cell proliferation, adhesion, motility, and invasion capacities were assessed in parental and transfected OVCAR3/CHP2 cell lines to explore the possible functions of CHP2 in ovarian cancer progression. RESULTS: With RT-PCR analysis, CHP2-transfected OVCAR3/CHP2 cancer cells showed high CHP2 gene expression, whereas non-transfected clones did not produce detectable CHP2 mRNA. CHP2-transfected OVCAR3/CHP2 cells showed increased proliferation rates and exhibited increased activities of cell adhesion, migration and invasion. The current study provides the first evidence that overexpression of the CHP2 gene affects the biological behavior of ovarian cancer cell line OVCAR3 and is one of key mechanisms for ovarian carcinoma progression, suggesting that CHP2 may be an attractive target for biological anticancer therapy.