Nonstructural N- and C-tails of Dbp2 confer the protein full helicase activities.

Human DDX5 and its yeast ortholog Dbp2 are ATP-dependent RNA helicases that play a key role in normal cell processes, cancer development, and viral infection. The crystal structure of the RecA1-like domain of DDX5 is available but the global structure of DDX5/Dbp2 subfamily proteins remains to be elucidated. Here, we report the first X-ray crystal structures of the Dbp2 helicase core alone and in complex with ADP at 3.22 Å and 3.05 Å resolutions, respectively. The structures of the ADP-bound post-hydrolysis state and apo-state demonstrate the conformational changes that occur when the nucleotides are released. Our results showed that the helicase core of Dbp2 shifted between open and closed conformation in solution but the unwinding activity was hindered when the helicase core was restricted to a single conformation. A small-angle X-ray scattering experiment showed that the disordered amino (N) tail and carboxy (C) tails are flexible in solution. Truncation mutations confirmed that the terminal tails were critical for the nucleic acid binding, ATPase, and unwinding activities, with the C-tail being exclusively responsible for the annealing activity. Furthermore, we labeled the terminal tails to observe the conformational changes between the disordered tails and the helicase core upon binding nucleic acid substrates. Specifically, we found that the nonstructural terminal tails bind to RNA substrates and tether them to the helicase core domain, thereby conferring full helicase activities to the Dbp2 protein. This distinct structural characteristic provides new insight into the mechanism of DEAD-box RNA helicases.

Improvement of vaginal probiotics Lactobacillus crispatus on intrauterine adhesion in mice model and in clinical practice.

BACKGROUND: Intrauterine adhesion (IUA) is a frequent acquired endometrial condition, for which there is no effective preventive or treatment. Previous studies have found that vaginal microbiota dysregulation is closely related to endometrial fibrosis and IUA. Therefore, we wondered whether restoration of vaginal microbiota by vaginal administration of L. crispatus could prevent endometrial fibrosis and ameliorate IUA. RESULTS: First, we created a mechanically injured mouse model of IUA and restored the mice's vaginal microbiota by the addition of L. crispatus convolvulus. The observations suggested that intrauterine injections of L. crispatus significantly decreased the degree of uterine fibrosis, the levels of IL-1ß and TNF-α in blood, and downregulated the TGF-ß1/SMADs signaling pathway in IUA mice. A therapy with L. crispatus considerably raised the abundance of the helpful bacteria Lactobacillus and Oscillospira and restored the balance of the vaginal microbiota in IUA mice, according to high-throughput sequencing. Then we conducted a randomized controlled trial to compare the therapeutic effect of L. crispatus with estrogen after transcervical resection of adhesion (TCRA). And the results showed that vaginal probiotics had a better potential to prevent intrauterine adhesion than estrogen. CONCLUSIONS: This study confirmed that L. crispatus could restore vaginal microbiota after intrauterine surgery, inhibit endometrial fibrosis, and finally play a preventive and therapeutic role in IUA. At the same time, it is a new exploration for the treatment of gynecological diseases with vaginal probiotics. CLINICAL TRIAL REGISTRATION: http://www.chictr.org.cn/ , identifier (ChiCTR1900022522), registration time: 15/04/2019.

Vitamin K3 Suppresses Pyroptosis in THP-1 Cells through Inhibition of NF-κB and JNK Signaling Pathways.

Vitamin K, a necessary nutritional supplement for human, has been found to exhibit anti-inflammatory activity. In the present study, we investigated the effects of vitamin K family on lipopolysaccharide (LPS) plus nigericin induced pyroptosis and explored the underlying mechanism of its action in THP-1 monocytes. Results showed that vitamin K3 treatment significantly suppressed THP-1 pyroptosis, but not vitamin K1 or K2, as evidenced by increased cell viability, reduced cellular lactate dehydrogenase (LDH) release and improved cell morphology. Vitamin K3 inhibited NLRP3 expression, caspase-1 activation, GSDMD cleavage and interleukin (IL)-1ß secretion in pyrophoric THP-1 cells. In addition, vitamin K3 inhibited the pro-inflammatory signaling pathways including nuclear factor-κB (NF-κB) and c-Jun N-terminal kinase (JNK). Vitamin K3 treatment also attenuated tissue damage and reduced serum LDH, IL-1ß and IL-6 levels in LPS-induced systemic inflammation of mice. The reduced myeloperoxidase (MPO) activityand F4/80 expression indicated that vitamin K3 effectively reduced the infiltration of neutrophils and macrophages. Moreover, NLRP3 expression in monocytes/macrophages were also decreased in vitamin K3-treatedmice after LPS challenge. These findings suggest that vitamin K3 potently alleviates systemic inflammation and organ injury via inhibition of pyroptosis in monocytes and may serve as a novel therapeutic strategy for patients with inflammatory diseases.

Resistance to Bacillus thuringiensis Cry1Ac toxin requires mutations in two Plutella xylostella ATP-binding cassette transporter paralogs.

The diamondback moth, Plutella xylostella, is a cosmopolitan pest and the first species to develop field resistance to toxins from the gram-positive bacterium Bacillus thuringiensis (Bt). Although previous work has suggested that mutations of ATP-binding cassette transporter subfamily C2 (ABCC2) or C3 (ABCC3) genes can confer Cry1Ac resistance, here we reveal that P. xylostella requires combined mutations in both PxABCC2 and PxABCC3 to achieve high-level Cry1Ac resistance, rather than simply a mutation of either gene. We identified natural mutations of PxABCC2 and PxABCC3 that concurrently occurred in a Cry1Ac-resistant strain (Cry1S1000) of P. xylostella, with a mutation (RA2) causing the mis-splicing of PxABCC2 and another mutation (RA3) leading to the premature termination of PxABCC3. Genetic linkage analysis showed that RA2 and RA3 were tightly linked to Cry1Ac resistance. Introgression of RA2 and RA3 enabled a susceptible strain (G88) of P. xylostella to obtain high resistance to Cry1Ac, confirming that these genes confer resistance. To further support the role of PxABCC2 and PxABCC3 in Cry1Ac resistance, frameshift mutations were introduced into PxABCC2 and PxABCC3 singly and in combination in the G88 strain with CRISPR/Cas9 mediated mutagenesis. Bioassays of CRISPR-based mutant strains, plus genetic complementation tests, demonstrated that the deletion of PxABCC2 or PxABCC3 alone provided < 4-fold tolerance to Cry1Ac, while disruption of both genes together conferred >8,000-fold resistance to Cry1Ac, suggesting the redundant/complementary roles of PxABCC2 and PxABCC3. This work advances our understanding of Bt resistance in P. xylostella by demonstrating mutations within both PxABCC2 and PxABCC3 genes are required for high-level Cry1Ac resistance.

Rapid diagnosis of seven high-risk human papillomavirus subtypes by a novel loop-mediated isothermal amplification method.

Current human papillomavirus (HPV) detection methods require complex instruments, skilled staff and have a high cost. Therefore, novel testing approaches are needed which are easy to implement, highly sensitive, and low cost. Loop-mediated isothermal amplification (LAMP) is an isothermal amplification technique. In this study, according to the conditions in China, a novel LAMP method for detecting seven high-risk HPV subtypes (16, 18, 33, 39, 45, 52, and 58) was designed and evaluated. The DNA from plasmid and cervical specimens was extracted using Chelex 100 and measured by qPCR and LAMP assay. LAMP products were observed under ultraviolet light. HPV sequences were successfully amplified and a plateau time of 19-75 min was maintained. The concentration of positive reactions ranged between 20 copies/µL and 200000 copies/µL. Additionally, there was no cross-reactivity between HPV16, 18, 33, 39, 45, 52, 58, 31, 35, 45, 51, 56, 59, 66, or 68. For clinical samples, the LAMP assay had high sensitivity and specificity for HPV16, 18, 33, 39, 45, 52, and 58. However, 5% (72/1447) of the samples tested yielded false-positive results. In conclusion, the novel LAMP assay for HPV16, 18, 33, 39, 45, 52, and 58 has high sensitivity and specificity, a low cost, and is simple and rapid to perform. The LAMP assay can improve HPV detection in resource-limited settings, especially in primary care hospitals and rural areas.

Correlation study on serum miR-222-3p and glucose and lipid metabolism in patients with polycystic ovary syndrome.

OBJECTIVE: microRNAs (miRNAs) play pivotal roles in polycystic ovary syndrome (PCOS), an endocrine and metabolic disorder that commonly occurs in women of childbearing age. This paper aimed to measure miR-222-3p expression in sera of PCOS patients and to explore its clinical value on PCOS diagnosis and prediction of diabetic and cardiovascular complications. METHODS: Totally 111 PCOS patients and 94 healthy people were recruited and assigned to the overweight (ow) group and non-overweight (non-ow) group, followed by determination of serum miR-222-3p expression. The diagnostic efficiency of miR-222-3p on PCOS ow and non-ow patients was analyzed. Correlations between miR-222-3p and glycolipid metabolic indicators and diabetic and cardiovascular complications in PCOS were analyzed. The downstream target of miR-222-3p was predicted and their binding relationship was verified. The correlation between PGC-1α and miR-222-3p was analyzed. RESULTS: miR-222-3p was highly-expressed in PCOS patients (p < 0.001), in especially PCOS ow patients. The area under the curve (AUC) of miR-222-3p diagnosing PCOS non-ow patients was 0.9474 and cut-off value was 1.290 (89.06% sensitivity, 98.11% specificity), indicating that non-ow people with serum miR-222-3p > 1.290 could basically be diagnosed with PCOS. AUC of miR-222-3p diagnosing PCOS ow patients was 0.9647 and cut-off value was 2.425 (85.11% sensitivity, 100% specificity), suggesting that ow people with serum miR-222-3p > 2.425 could basically be diagnosed with PCOS. miR-222-3p was positively-correlated with fasting plasma glucose (FPG), fasting insulin (FINS), homeostatic model assessment-insulin resistance (HOMA-IR), and low-density lipoprotein cholesterol (LDL-C) and negatively-correlated with high-density lipoprotein cholesterol (HDL-C). miR-222-3p was independently-correlated with diabetic and cardiovascular complications in PCOS (p < 0.05). High expression of miR-222-3p predicted high risks of diabetic and cardiovascular complications in PCOS. miR-222-3p targeted PGC-1α and was negatively-correlated with PGC-1α (r = - 0.2851, p = 0.0224; r = - 0.3151, p = 0.0310). CONCLUSION: High expression of miR-222-3p assisted PCOS diagnosis and predicted increased risks of diabetic and cardiovascular complications. miR-222-3p targeted PGC-1α and was negatively-correlated with PGC-1α.

Increased functional interaction within frontoparietal network during working memory task in major depressive disorder.

Abnormal fronto-parietal activation has been suggested as a neural underpinning of the working memory (WM) deficits in major depressive disorder (MDD). However, the potential interaction within the frontoparietal network during WM processing in MDD remains unclear. This study aimed to examine the role of abnormal functional interactions within frontoparietal network in the neuropathological mechanisms of WM deficits in MDD. A total of 40 MDD patients and 47 demographic matched healthy controls (HCs) were included. Functional magnetic resonance imaging and behavioral data were collected during numeric n-back tasks. The psychophysiological interaction and dynamic causal modelling methods were applied to investigate the connectivity within the frontoparietal network in MDD during n-back tasks. The psychophysiological interaction analysis revealed that MDD patients showed increased functional connectivity between the right inferior parietal lobule (IPL) and the right dorsolateral prefrontal cortex (dlPFC) compared with HCs during the 2-back task. The dynamic causal modelling analysis revealed that MDD patients had significantly increased forward modulation connectivity from the right IPL to the right dlPFC than HCs during the 2-back task. Partial correlation was used to calculate the relationship between connective parameters and psychological variables in the MDD group, which showed that the effective connectivity from right IPL to right dlPFC was correlated negatively with the sensitivity index d' of WM performances and positively with the depressive severity in MDD group. In conclusion, the abnormal functional and effective connectivity between frontal and parietal regions might contribute to explain the neuropathological mechanism of working memory deficits in major depressive disorder.

Insecure attachment and maladaptive emotion regulation mediating the relationship between childhood trauma and borderline personality features.

BACKGROUND: Previous studies suggested that childhood trauma is an important etiologic factor for the development of borderline personality disorder (BPD). Moreover, insecure attachment and maladaptive emotion regulation (ER) might be related to childhood trauma and BPD. This study was aimed to explore the relationships among childhood trauma, insecure attachment, maladaptive ER, and BPD features. METHODS: A cohort of 637 patients with psychological disorders completed a series of psychometric instruments such as the Personality Diagnostic Questionnaire-4+ (PDQ-4+), the 23-Item Borderline Symptom List, the Childhood Trauma Questionnaire, the Attachment Style Questionnaire, and the Cognitive Emotion Regulation Questionnaire. The path analyses were conducted to investigate the experience-driven model that whether insecure attachment and maladaptive ER could mediate the relationship between childhood trauma and BPD features. The random forest regression was performed to select variables that contribute significantly to BPD features, which variables would be incorporated into the data-driven model to further confirm the experience-driven model. RESULTS: Both the experience-driven model and the data-driven model verified that there were three significant mediation pathways (childhood trauma → insecure attachment/maladaptive ER → BPD features, childhood trauma → insecure attachment → maladaptive ER → BPD features; all p < .05), and the most weighted mediation pathway by which childhood trauma influencing the BPD features was through insecure attachment and then through maladaptive ER (weighted 53.16%). CONCLUSION: The influence of childhood trauma on BPD features was mainly mediated by the combination of insecure attachment and maladaptive emotion regulation.

Evaluation of EZH2 expression, BRAF V600E mutation, and CDKN2A/B deletions in epithelioid glioblastoma and anaplastic pleomorphic xanthoastrocytoma.

INTRODUCTION: Epithelioid glioblastoma (EGBM) and anaplastic pleomorphic xanthoastrocytoma (APXA) are two rare entities with different prognoses. However, they share certain morphological and molecular features. MATERIALS AND METHODS: To better recognize EGBM and APXA and identify the prognostic factors associated with these tumors, EZH2 status, BRAF V600E mutations, and CDKN2A/B deletions were assessed in 15 APXA and 13 EGBM cases. RESULTS: The expression level of EZH2 was found to increase with tumor grade. Overexpression of EZH2 occurred in 69.2% (9/13) of EGBM cases and 20% (3/15) of APXA cases. In addition, 72.7% (8/11) of EGBM and 12.5% (1/8) of APXA cases harbored a CDKN2A homozygous deletion based on fluorescence in situ hybridization. BRAF V600E mutations were detected in 80% (8/10) of EGBM cases and 42.9% (3/7) of APXA cases. Furthermore, EGBM, which exhibited co-existing low-grade glioma-like lesions, was found to have strong EZH2 expression and high Ki-67 indexes only in epithelioid cells and not in low grade lesions. Univariate analysis demonstrated that abundant epithelioid cells, extensive necrosis, EZH2 overexpression and BRAF V600E mutations were significantly associated with decreased overall survival in EGBM and APXA patients (P < 0.05). CONCLUSIONS: The results suggested that testing for EZH2 expression and BRAF V600E mutations might be helpful to evaluate the prognoses of EGBM and APXA patients. The presence of heterogeneous EZH2 expression in biphasic EGBMs could also contribute to malignant progression.

Clinical Significance of Transcription Factor 7 (TCF7) as a Prognostic Factor in Gastric Cancer.

BACKGROUND Transcription factor 7 (TCF7) plays an essential role in Wnt signaling by interacting with ß-catenin. Emerging evidence demonstrates that overexpression of TCF7 promotes progression or correlates with poor progression in several types of cancers, but the functions of TCF7 in gastric cancer (GC) have not been revealed. MATERIAL AND METHODS A total of 168 patients with GC who underwent radical surgeries were collected and regarded as the test cohort. The expression of TCF7 in the 168 patients was detected with immunohistochemistry. Moreover, the mRNA levels of TCF7 in 11 pairs of GC and adjacent tissues were detected with quantitative real-time PCR (qRT-PCR). The correlations between TCF7 and the clinicopathological factors were evaluated with the chi-square test, and the prognostic value of TCF7 in GC was investigated with univariate analysis and multivariate analysis. RESULTS The mRNA levels of TCF7 in GC tissues were significantly higher than in corresponding tumor adjacent tissues. The patients of low TCF7 expression and high TCF7 expression accounted for 76.79% (129/168) and 23.21% (39/168), respectively. In our experiments, TCF7 was significantly associated with positive lymphatic invasion (P=0.022) and metastasis (P<0.001). The high expression of TCF7 was correlated with low survival rates (P=0.012) and was confirmed as an independent prognostic factor (HR=1.92, 95%CI =1.06-3.47, P=0.031) of GC in multivariate analysis. CONCLUSIONS TCF7 expression is correlated with metastasis and is an independent prognostic factor of GC. TCF7 detection of GC could help stratify the patients with high risk and guide precise treatment.

Long non-coding RNA MIAT promotes growth and metastasis of colorectal cancer cells through regulation of miR-132/Derlin-1 pathway.

OBJECTIVE: Recently, long non-coding RNA (lncRNA) MIAT has been demonstrated as an oncogenic gene in several types of cancer. However, the role and mechanism of MIAT in colorectal cancer (CRC) have not been investigated. METHODS: Real-time PCR was used to measure MIAT expression in CRC tissues and cells. Small interfering RNA specific for MIAT (si-MIAT) was used to down-regulate MIAT expression in CRC cells. The interaction of MIAT and miR-132 was measured by RNA pull-down assay. The effect of si-MIAT on CRC cells apoptosis and metastasis were measured by flow cytometry assay, invasion and migration assay, respectively. RESULTS: In present study, we found that MIAT was highly expressed in CRC tissues and cells. MIAT knockdown inhibited proliferation, migration and invasion and enhanced apoptosis of CRC cells. Further, we demonstrated that MIAT acted as a competing endogenous RNA for miR-132, antagonized its functions, and resulted in the de-repression of its target gene Derlin-1, which acted as an oncogene in promoting growth and metastasis of CRC cells. In LOVO and SW480 cells with si-MIAT, miR-132 inhibitor resulted in an increase of cell proliferation, migration and invasion and a decrease of cell apoptosis, which was partially abolished by transfection of Derlin-1 shRNA. CONCLUSIONS: Our data indicated that highly expressed MIAT was an oncogenic lncRNA that promoted the growth and metastasis of CRC through miR-132/Derlin-1 axis.

Identification of transcripts involved in digestion, detoxification and immune response from transcriptome of Empoasca vitis (Hemiptera: Cicadellidae) nymphs.

Tea production has been significantly impacted by the false-eye leafhopper, Empoasca vitis (Göthe), around Asia. To identify the key genes which are responsible for nutrition absorption, xenobiotic metabolism and immune response, the transcriptome of either alimentary tracts or bodies minus alimentary tract of E. vitis was sequenced and analyzed. Over 31 million reads were obtained from Illumina sequencing. De novo sequence assembly resulted in 52,182 unigenes with a mean size of 848nt. The assembled unigenes were then annotated using various databases. Transcripts of at least 566 digestion-, 224 detoxification-, and 288 immune-related putative genes in E. vitis were identified. In addition, relative expression of highly abundant transcripts was verified through quantitative real-time PCR. Results from this investigation provide genomic information about E. vitis, which will be helpful in further study of E. vitis biology and in the development of novel strategies to control this devastating pest.

Land surface temperature retrieval from AMSR-E passive microwave data.

The land surface temperature (LST) is a key parameter for energy balance, evapotranspiration and climate change. In this study, two new methods of LST retrieval from passive microwave data are developed: one is deriving LST only using single-channel dual-polarized data based on the relationship between the emissivity and microwave polarization difference index (MPDI) (denoted as Method 1); the other one is deriving LST using the traditional multi-channel method with prior knowledge of the normalized difference vegetation index (NDVI) (denoted as Method 3). Taking Moderate Resolution Imaging Spectroradiometer (MODIS) LST products as the actual LSTs, the coefficients for these algorithms are determined. From the results for the year 2008, it is demonstrated that the root mean square errors (RMSEs) for the LST retrieval using Method 3 are the smallest and range from 2.92 K to 3.44 K, the RMSEs for the LST retrieval using traditional multi-channel method (denoted as Method 2) range from 3.07 K to 4.05 K, and the worst results come from Method 1, whose RMSEs range from 3.11 K to 4.13 K at a frequency of 89 GHz. This could be caused by the fact that the NDVI provides substantial emissivity knowledge in Method 3, and much richer vegetation could result in a more accurate emissivity estimation.

A clinicopathologic study of extraventricular neurocytoma.

In 2007, extraventricular neurocytoma was classified as a separate entity among glioneuronal tumors. However, extraventricular neurocytoma is not fully understood and may be misdiagnosed. Here, we describe the clinical and pathological features, prognoses, and treatments of 13 extraventricular neurocytoma cases, and compare their immunophenotypes with those of oligodendroglioma, diffuse astrocytoma, and ependymoma. Six typical and 7 atypical cases comprised the 13 extraventricular neurocytoma cases. Histological features included oligodendroglioma-like perinuclear halo, neuropil-like matrix, ganglion or ganglioid cells, perivascular pseudorosettes, vessel hyalinization, calcifications, and myxoid degeneration. Atypical histological features included increased mitotic figures, focal necrosis, endothelial cell proliferation, and/or a Ki-67 index of >2%. All lesions expressed synaptophysin and microtubule-associated protein-2, which distinguished them from other similar tumors. Two patients with atypical extraventricular neurocytoma had tumor recurrence, one of whom had cerebrospinal fluid dissemination, suggesting that atypical histological features might represent adverse prognostic factors. In conclusion, the present study identified morphological and immunohistochemical features that would aid the differential diagnosis of extraventricular neurocytoma. In addition, radiotherapy with subtotal resection could be considered an effective treatment for extraventricular neurocytoma, but because a pediatric patient died of intracranial hemorrhage during radiotherapy, radiotherapy-related side effects should be considered, especially when treating children. Additional cases with long-term follow-up are needed to develop optimal management protocols for extraventricular neurocytoma.

The clinicopathological features of pituicytoma and the differential diagnosis of sellar glioma.

Pituicytoma is rare and difficult to diagnose. This study explored the clinicopathological features, immunophenotype and differential diagnosis of pituicytoma. We compared 11 cases of pituicytoma and 26 cases of sellar glioma (16 pilocytic astrocytomas, four diffuse astrocytomas, three pilomyxoid astrocytomas, and three third ventricle chordoid gliomas). The 11 pituicytoma cases involved six men and five women (age: 33-65 years). Three of the 11 patients experienced recurrence due to a residual tumor, and one patient underwent three surgeries during a 6-month period. Imaging findings revealed tumors were in the intrasellar region (four cases), suprasellar region (four cases) and intra-suprasellar regions (three cases). The tumor diameters were 1.3-3.8 cm, and the preoperative diagnoses were pituitary adenoma, craniopharyngioma and meningioma. The tumors were solid and contained spindle or slightly chubby cells that were densely arranged with visible cleft-like or expanded sinusoid structures. The cells had vague boundaries, circular nuclei, fine chromatin, and a small nucleolus. Immunohistochemical staining of the pituicytomas revealed positive expression of thyroid transcription factor-1 (TTF-1) and S-100 protein (S-100), positive focal expression of glial GFAP;(five of 11 cases), and negative oligodendrocyte transcription factor 2 (Olig2), CD34 and neurofilament expression. The Ki67 index was 6% in one case and 1-2% in the other cases. Unlike pituicytoma, most sellar glioma cases exhibited GFAP and Olig2 expression, and negative TTF-1 expression. Third ventricle chordoid gliomas expressed TTF-1, GFAP and CD34, and were negative for Olig2. Our results indicate that pituicytoma typically involves dense arrangements of spindle or slightly chubby cells. The morphology is occasionally atypical, with ependymoma-like or meningioma-like structures, and occasionally exhibits pilomyxoid degeneration. Abundant sinusoids are characteristic of hemorrhagic tumors. The dense spindle cell arrangement is a relatively specific morphology, and staining for GFAP, TTF-1, Olig2 and CD4 may help differentiate pituicytoma from sellar glioma.

Overexpression of Notch3 and pS6 Is Associated with Poor Prognosis in Human Ovarian Epithelial Cancer.

Notch3 and pS6 play important roles in tumor angiogenesis. To assess the expression of Notch3 and pS6 in Chinese ovarian epithelial cancer patients, a ten-year follow-up study was performed in ovarian epithelial cancer tissues from 120 specimens of human ovarian epithelial cancer, 30 specimens from benign ovarian tumors, and 30 samples from healthy ovaries by immunohistochemistry. The results indicate that the expression of Notch3 and pS6 was higher in ovarian epithelial cancer than in normal ovary tissues and in benign ovarian tumor tissues (p < 0.01). In tumor tissues, Notch3 expression and pS6 expression were negatively associated with age (p > 0.05) but positively associated with clinical stage, pathological grading, histologic type, lymph node metastasis, and ascites (p < 0.05 or p < 0.01). A follow-up survey of 64 patients with ovarian epithelial cancer showed that patients with high Notch3 and pS6 expression had a shorter survival time (p < 0.01), in which the clinical stage (p < 0.05) and Notch3 expression (p < 0.01) played important roles. In conclusion, Notch3 and pS6 are significantly related to ovarian epithelial cancer development and prognosis, and their combination represents a potential biomarker and therapeutic target in ovarian tumor angiogenesis.

[Clinicopathologic features of embryonal tumor with multilayered rosettes and gene analysis on chromosome 19q13.42].

OBJECTIVE: To investigate the clinicopathologic features and 19q13.42 gene changes in embryonal tumors with multilayered rosettes (ETMR). METHODS: Immunohistochemistry and fluorescence in situ hybridization (FISH) were performed in three ETMRs. RESULTS: The average age of the patients were 34 months. Imaging revealed huge masses with inhomogeneous enhancement and two cases showed cystic lesions. Follow-up data showed 14 and 38 months survival in two children, the third had a recurrence 4 months after operation. Morphologically, the tumor was mainly composed of dense small primitive neuroepithelial cells in patchy or multilayer rosettes within a background of advanced neuronal differentiation, containing neurocytes, ganglion cells, and neuropil-like background. Immunohistochemical staining showed the neuronal marker, synaptophysin, was positive in differentiated areas. Nestin as a neural stem cell marker was immunoreactive in the primitive neuroepithelial cells including multilayered rosettes. Neurons with positive expression of NeuN were observed occasionally. Ki-67 index was up to 40%-80% in the undifferentiated cells and rosettes, but was only 1%-3% in the differentiated areas. CD99 was positive in perivascular papillary pattern areas in one case. 19q13.42 amplification was detected in more than 30% of tumor cells in all cases. CONCLUSIONS: ETMR is a unique entity with distinctive clinical and pathological features. Chromosome 19q13.42 abnormality is valuable for confirming the diagnosis and for further treatment research.

Xanthone glycosides from Swertia bimaculata with α-glucosidase inhibitory activity.

Seven new xanthone glycosides (1-7) were isolated from the n-butanol extract of Swertia bimaculata, together with six known compounds (8-13). Their structures were elucidated on the basis of extensive spectroscopic analyses (1D- and 2D-NMR, HRESIMS, UV, and IR) and comparison with data reported in the literature. All the compounds were evaluated for their α-glucosidase inhibitory activities in vitro, and compounds 3, 4, and 7 exhibited significant activities to inhibit α-glucosidase. Meanwhile the effects of different substitutions on the α-glucosidase inhibitory activity of xanthone glycosides from S. bimaculata are also discussed.

[Study on Quality Standard for Panax japonicus Rhizome].

OBJECTIVE: To establish a quality standard for Panax japonicus rhizome. METHODS: Ginsenoside Ro and Chikusetsusaponin IVa were used as reference substances in the TLC identification and HPLC method. Additionally, acid insoluble ash and moisture were determined according to the procedures recorded in the Appendix of Chinese Pharmacopeia (2010 edition). RESULTS: The TLC identification with GF, showed a good resolution with clear spots and its optimum developer was the underlayer of chloroform-methanol-formic acid-water = 4.5:1.5: 0.1:0.3. The content of Ginsenoside Ro and Chikusetsusaponin N a were determined by HPLC. The mixture of acetonitrile and water (0.15% phosphate) with gradient elution as the mobile phase was used at a flow rate of 1.0 mL/min, the detection wavelength at 203 nm and the column temperature at 40 °C. The calibration curve was linear in the range of 31.25-2,000 g/mL for Ginsenoside Ro and Chikusetsusaponin IVa (r = 0.9999, respectively). The average recovery was 101.19% and 102.50%, and RSD was 1.59% and 1.80% respectively. The content of Ginsenoside Ro and Chikusetsusaponin IVa was no less than 1.5% respectively. An average content of moisture was 7.36% and acid insoluble ash was 0.84%. CONCLUSION: These methods are producible, sensitive and simple, which can be used to control the quality of Panax japonicus rhizome.

Vaginal microbiota: Potential targets for vulvovaginal candidiasis infection.

Vulvovaginal candidiasis (VVC) is the second most common cause of vaginal infection globally after bacterial vaginosis (BV) and associated with adverse reproductive and obstetric outcomes, including preterm delivery, sexually transmitted infections and pelvic inflammatory disease. Although effective control of VVC is achievable with the use of traditional treatment strategies (i.e., antifungals), the possibility of drug intolerance, treatment failure and recurrence, as well as the appearance of antifungal-resistant Candida species remain critical challenges. Therefore, alternative therapeutic strategies against VVC are urgently required. In recent years, an improved understanding of the dysbiotic vaginal microbiota (VMB) during VVC has prompted the consideration of administering -biotics to restore the balance of the VMB within the context of VVC prevention and treatment. Here, we aim to summarize the current evidence of the anti-Candida effects of probiotics, postbiotics and synbiotics and their potential use as an alternative/complementary therapy against VVC. Additionally, this review discusses advantages and challenges associated with the application of -biotics in VVC to provide guidance for their later use. We also review new developments in VVC therapy, i.e., vaginal microbiota transplantation (VMT) as an emerging live biotherapeutic therapy against VVC and discuss existing shortcomings associated with this nascent field, expecting to stimulate further investigations for introduction of new therapies against VVC.