PKMYT1 aggravates the progression of ovarian cancer by targeting SIRT3.

OBJECTIVE: This experiment aims to elucidate the role of PKMYT1 in the malignant progression of ovarian cancer (OC) and its underlying mechanism. PATIENTS AND METHODS: Expression pattern of PKMYT1 in 43 paired OC tissues and adjacent normal ones was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The potential relationship between PKMYT1 level and clinical data of OC patients was analyzed. PKMYT1 level in OC patients either with distant metastasis or not was examined. Through Cell Counting Kit (CCK-8) and transwell assay, influences of PKMYT1 on proliferative and metastatic abilities in 3AO and CAOV3 cells were assessed. At last, the role of PKMYT1/SIRT3 regulatory loop in the progression of OC was identified. RESULTS: PKMYT1 was upregulated in OC tissues relative to controls. OC patients accompanied with distant metastasis had higher abundance of PKMYT1. High level of PKMYT1 predicted worse prognosis in OC patients. Knockdown of PKMYT1 attenuated proliferative, migratory, and invasive abilities in OC cells. Moreover, SIRT3 was downregulated in OC tissues, which was negatively correlated to PKMYT1. Silencing of SIRT3 could abolish the regulatory effect of PKMYT1 on proliferative and metastatic abilities in OC. CONCLUSIONS: Upregulated PKMYT1 in OC is closely linked to distant metastasis and poor prognosis. PKMYT1 accelerates the malignant progression of OC via negatively regulating SIRT3.

Effectiveness of second-dose varicella vaccination as post-exposure prophylaxis: a prospective cohort study.

OBJECTIVES: Although administration of a second dose of varicella vaccine (2nd-dose VarV) to individuals who have previously received one-dose VarV has been recommended as a post-exposure prophylaxis (PEP) strategy for outbreak control, the effectiveness of this strategy remains unclear. We evaluated the vaccine effectiveness (VE) of 2nd-dose VarV as PEP among students involved in 129 varicella outbreaks in Shanghai, China from 2013 to 2016. METHODS: Students who had received one-dose VarV more than 5 years prior to varicella exposure were eligible to receive 2nd-dose VarV as PEP. We evaluated the VE using the following formula: VE = (1 - hazard ratio (HR)) × 100%. RESULTS: A total of 6762 students were eligible for 2nd-dose VarV, of whom 58.6% accepted PEP after varicella exposure. The adjusted VE of 2nd-dose VarV as PEP was 77% (95% confidence interval 64-85%). In addition, the adjusted VE of 2nd-dose VarV as PEP in affected classrooms with high vaccine uptake was higher than that in classrooms with lower vaccine uptake (87% vs. 69%). The adjusted VE was also higher in students who received 2nd-dose VarV within 3 days of exposure than those who received it more than 3 days post exposure (77% vs. 64%). CONCLUSIONS: These Results suggest that administration of 2nd-dose VarV as PEP is an appropriate intervention for outbreak control in countries where two-dose VarV has not been adopted.

Assessments of M-mode color echocardiography on fetal right ventricular diastolic function with umbilical cord around neck.

OBJECTIVE: To investigate the fetal right ventricular diastolic function under the condition of umbilical cord around neck (UCAN), and analyze the changes of the right ventricular propagation velocity (Vp), then discuss the clinical value of the color M-mode echocardiography in the evaluation of fetal ventricular diastolic function quantitatively. PATIENTS AND METHODS: All patients enrolled were with singleton pregnancy from Cangzhou Central Hospital from December 2013 to December 2015 as the experimental group. The control group consisted of normal fetuses without UCAN and the experimental group consisted of the fetuses with UCAN. Besides, this paper analyzed values of Tei index of the left and right ventricle as well as Vp of the right ventricle diastole using color M-mode echocardiography. RESULTS: The Vp values of the experimental group were significantly lower than those of the control group (p < 0.05); the Tei index of the right ventricle of the experimental group was significantly higher than that of the control group (p < 0.05); the Tei indexes of the left and right ventricles of the experimental group had no statistical difference (p > 0.05). The heart function and the right ventricular diastolic function were reduced in fetuses with UCAN; however, the effect of the left and the right ventricular diastolic function had no significant changes in fetuses with UCAN. CONCLUSIONS: It had great significance to select the appropriate index of cardiac function for estimating the right ventricular diastolic function and the whole heart function of UCAN, and it is of huge practical application value in clinical practice.

[Individual dose monitoring and health status of radiation workers in some Three A hospitals of Zhejiang province].

Objective: To investigate the individual irridiateddose levelandhealth statusofoccupational externalexposureamong radiationworkers inThreeA hospitals ofZhejiangprovince,andprovideevidence for occupationalhealthmanagement. Methods: 367 different typesof radiationworkerswereexperienced thedose monitoring and health examination from January toDecember, 2015, according to the requirements of"SpecificationsofIndividualMonitoring forOccupationalExternalExposure"and"Specifications forOccupational healthsurveillance forradiationworkers".The resultsofdosemonitoring,chromosomeaberration rate, lensopacity rate,hemogramand thyroid functionwerestatisticallyanalyzed. Results: Theannualeffectivedoseamong radiation workers fromZhejiangprovince in2015was0.13mSv,98.91%of them less than1mSv,whichunder the limit standardofstate(20mSv/a).Thechromosomeaberration(dicentric)detection rateswere7.41%and4.35%, from nuclearmedicinegroupand interventionalgroup respectively,whichhigher thandiagnostic radiologygroup, the differencewasstatisticallysignificant(χ(2)=13.686,8.092,P<0.01).Besides,1caseofsuspiciouschronic radiation dermatitiswas found in the interventiongroup.Radiation lengthhadsignificanteffecton lensopacity rate(P<0.01), lensopacity increasedwith the increasing lengthof the linear trend (χ(2)trend=16.363,P<0.01),and the incidence ofabnormalthyroid function theabnormalrateof lymphocyte ratiohad significantdifferenceamong theagegroups (P<0.05). Conclusion: Although, theoccupationalexternalexposureamong radiationworkersinThreeAhospitals ofZhejiangprovince issafe, long-term lowdosesofionizing radiation stillhascertainhealtheffectson the fieldof nuclearmedicineand interventionalradiologystaff,suchaseye lens,cytogenetics,nailsandskin.

SMARCA4/Brg1 coordinates genetic and epigenetic networks underlying Shh-type medulloblastoma development.

Recent large-scale genomic studies have classified medulloblastoma into four subtypes: Wnt, Shh, Group 3 and Group 4. Each is characterized by specific mutations and distinct epigenetic states. Previously, we showed that a chromatin regulator SMARCA4/Brg1 is required for Gli-mediated transcription activation in Sonic hedgehog (Shh) signaling. We report here that Brg1 controls a transcriptional program that specifically regulates Shh-type medulloblastoma growth. Using a mouse model of Shh-type medulloblastoma, we deleted Brg1 in precancerous progenitors and primary or transplanted tumors. Brg1 deletion significantly inhibited tumor formation and progression. Genome-wide expression analyses and binding experiments indicate that Brg1 specifically coordinates with key transcription factors including Gli1, Atoh1 and REST to regulate the expression of both oncogenes and tumor suppressors that are required for medulloblastoma identity and proliferation. Shh-type medulloblastoma displays distinct H3K27me3 properties. We demonstrate that Brg1 modulates activities of H3K27me3 modifiers to regulate the expression of medulloblastoma genes. Brg1-regulated pathways are conserved in human Shh-type medulloblastoma, and Brg1 is important for the growth of a human medulloblastoma cell line. Thus, Brg1 coordinates a genetic and epigenetic network that regulates the transcriptional program underlying the Shh-type medulloblastoma development.

Clustering autism: using neuroanatomical differences in 26 mouse models to gain insight into the heterogeneity.

Autism is a heritable disorder, with over 250 associated genes identified to date, yet no single gene accounts for >1-2% of cases. The clinical presentation, behavioural symptoms, imaging and histopathology findings are strikingly heterogeneous. A more complete understanding of autism can be obtained by examining multiple genetic or behavioural mouse models of autism using magnetic resonance imaging (MRI)-based neuroanatomical phenotyping. Twenty-six different mouse models were examined and the consistently found abnormal brain regions across models were parieto-temporal lobe, cerebellar cortex, frontal lobe, hypothalamus and striatum. These models separated into three distinct clusters, two of which can be linked to the under and over-connectivity found in autism. These clusters also identified previously unknown connections between Nrxn1α, En2 and Fmr1; Nlgn3, BTBR and Slc6A4; and also between X monosomy and Mecp2. With no single treatment for autism found, clustering autism using neuroanatomy and identifying these strong connections may prove to be a crucial step in predicting treatment response.

The pro-longevity gene FoxO3 is a direct target of the p53 tumor suppressor.

FoxO transcription factors have a conserved role in longevity, and act as tissue-specific tumor suppressors in mammals. Several nodes of interaction have been identified between FoxO transcription factors and p53, a major tumor suppressor in humans and mice. However, the extent and importance of the functional interaction between FoxO and p53 have not been fully explored. Here, we show that p53 regulates the expression of FoxO3, one of the four mammalian FoxO genes, in response to DNA damaging agents in both mouse embryonic fibroblasts and thymocytes. We find that p53 transactivates FoxO3 in cells by binding to a site in the second intron of the FoxO3 gene, a genomic region recently found to be associated with extreme longevity in humans. While FoxO3 is not necessary for p53-dependent cell cycle arrest, FoxO3 appears to modulate p53-dependent apoptosis. We also find that FoxO3 loss does not interact with p53 loss for tumor development in vivo, although the tumor spectrum of p53-deficient mice appears to be affected by FoxO3 loss. Our findings indicate that FoxO3 is a p53 target gene, and suggest that FoxO3 and p53 are part of a regulatory transcriptional network that may have an important role during aging and cancer.

Targeted resequencing of a genomic region influencing tameness and aggression reveals multiple signals of positive selection.

The identification of the causative genetic variants in quantitative trait loci (QTL) influencing phenotypic traits is challenging, especially in crosses between outbred strains. We have previously identified several QTL influencing tameness and aggression in a cross between two lines of wild-derived, outbred rats (Rattus norvegicus) selected for their behavior towards humans. Here, we use targeted sequence capture and massively parallel sequencing of all genes in the strongest QTL in the founder animals of the cross. We identify many novel sequence variants, several of which are potentially functionally relevant. The QTL contains several regions where either the tame or the aggressive founders contain no sequence variation, and two regions where alternative haplotypes are fixed between the founders. A re-analysis of the QTL signal showed that the causative site is likely to be fixed among the tame founder animals, but that several causative alleles may segregate among the aggressive founder animals. Using a formal test for the detection of positive selection, we find 10 putative positively selected regions, some of which are close to genes known to influence behavior. Together, these results show that the QTL is probably not caused by a single selected site, but may instead represent the joint effects of several sites that were targets of polygenic selection.

HSP-70 mitigates LPS/SKI-induced cell damage by increasing sphingosine kinase 1 (SK1).

Heat shock proteins (HSPs) are potent protectors of cellular integrity against environmental stresses, including toxic microbial products. To investigate the mechanism of HSP-70 cell protection against bacterial lipopolysaccharide (LPS), we established a stable HSP-70 gene-transfected RAW 264.7 murine macrophage model of LPS-induced cell death. Bacterial LPS increases the activity of sphingosine kinase 1 (SK1), which catalyzes formation of sphingosine-1-phosphate (S1P). S1P functions as a critical signal for initiation and maintenance of diverse aspects of immune cell activation and function. When mouse macrophages were incubated with Escherichia coli LPS (1 microg/ml) and sphingosine kinase inhibitor (SKI, 5 microM), 90% of cells died. Neither LPS nor SKI alone at these doses damaged the cells. The LPS/SKI-induced cell death was partially reversed by overexpression of HSP-70 in gene-transfected macrophages. The specificity of HSP-70 in this reversal was demonstrated by transfection of HSP-70-specific siRNA. Down-regulation of HSP-70 expression after transfection of siRNA specific for HSP-70 was associated with increased LPS/SKI-induced cell damage. Overexpression of human or murine HSP-70 (HSPA1A and Hspa1a, respectively) increased both cellular SK1 mRNA and protein levels. Cellular heat shock also increased SK1 protein. These studies confirm the importance of SK1 as a protective moiety in LPS-induced cell injury and demonstrate that HSP-70-mediated protection from cells treated with LPS/SKI is accompanied by upregulating expression of SK1. HSP-70-mediated increases in SK1 and consequent increased levels of S1P may also play a role in protection of cells from other processes that lead to programmed cell death.

TRED: a transcriptional regulatory element database, new entries and other development.

Transcriptional factors (TFs) and many of their target genes are involved in gene regulation at the level of transcription. To decipher gene regulatory networks (GRNs) we require a comprehensive and accurate knowledge of transcriptional regulatory elements. TRED (http://rulai.cshl.edu/TRED) was designed as a resource for gene regulation and function studies. It collects mammalian cis- and trans-regulatory elements together with experimental evidence. All the regulatory elements were mapped on to the assembled genomes. In this new release, we included a total of 36 TF families involved in cancer. Accordingly, the number of target promoters and genes for TF families has increased dramatically. There are 11,660 target genes (7479 in human, 2691 in mouse and 1490 in rat) and 14,908 target promoters (10,225 in human, 2985 in mouse and 1698 in rat). Additionally, we constructed GRNs for each TF family by connecting the TF-target gene pairs. Such interaction data between TFs and their target genes will assist detailed functional studies and help to obtain a panoramic view of the GRNs for cancer research.

A high efficiency cloning and expression system for proteomic analysis.

The recent description of the complete genomes of the two most pathogenic species of Brucella opens the way for genome-based analysis of the antigenicity of their proteins. In the present report, we describe a bench-level high-efficiency cloning and expression system (HECES) that allow expression of large numbers of Brucella proteins based on genomic sequence information. Purified proteins are produced with high efficiency in a microarray format conducive to analysis of their sero-reactivity against serum from immunized animals. This method is applicable at either small or large scale of protein processing. While it does not require robotics, the format is amenable to robotic implementation for all aspects of the process and subsequent analysis of protein characteristics. This method will allow selection of new reagents for diagnosis of brucellosis and development of vaccine against Brucella, an important zoonotic disease and biothreat agent.

Geldanamycin prevents hemorrhage-induced ATP loss by overexpressing inducible HSP70 and activating pyruvate dehydrogenase.

Hemorrhage in mice results in decreased ATP levels in the jejunum, lung, kidney, heart, and brain but not in liver tissue lysates, albeit at variable levels and time kinetics. The decreased protein expression and activity of pyruvate dehydrogenase (PDH) accounted for the hemorrhage-induced ATP loss. Treatment with geldanamycin (GA; 1 microg/g body wt), a known inducer of heat shock protein (HSP)70, inhibited the hemorrhage-induced ATP loss in the jejunum, lung, heart, kidney, and brain. GA was found to increase PDH protein, preserve PDH enzymatic activity, and inhibit mucosal injury in jejunum tissues. GA-induced HSP70i was found to form complexes with PDH protein. HSP70 gene transfer into intestinal epithelial cells promoted PDH and ATP levels, whereas HSP70 short interfering RNA limited them. We conclude that agents able to increase the expression of HSP70 and PDH may be of value in reducing pathology resulting from hemorrhage-associated ATP loss.

Cloning, expression, and purification of Brucella suis outer membrane proteins.

Brucella, an aerobic, nonsporeforming, nonmotile Gram-negative coccobacillus, is a NIH/CDC category B bioterror threat agent that causes incapacitating human illness. Medical defense against the bioterror threat posed by Brucella would be strengthened by development of a human vaccine and improved diagnostic tests. Central to advancement of these goals is discovery of bacterial constituents that are immunogenic or antigenic for humans. Outer membrane proteins (OMPs) are particularly attractive for this purpose. In this study, we cloned, expressed, and purified seven predicted OMPs of Brucella suis. The recombinant proteins were fused with 6-His and V5 epitope tags at their C termini to facilitate detection and purification. The B. suis surface genes were PCR synthesized based on their ORF sequences and directly cloned into an entry vector. The recombinant entry constructs were propagated in TOP 10 cells, recombined into a destination vector, pET-DEST42, then transformed into Escherichia coli BL21 cells for IPTG-induced protein expression. The expressed recombinant proteins were confirmed with Western blot analysis using anti-6-His antibody conjugated with alkaline phosphatase. These B. suis OMPs were captured and purified using a HisGrab plate. The purified recombinant proteins were examined for their binding activity with antiserum. Serum derived from a rabbit immunized intramuscularly with dialyzed cell lysate of Brucella rough mutant WRR51. The OMPs were screened using the rabbit antiserum and purified IgG. The results suggested that recombinant B. suis OMPs were successfully cloned, expressed and purified. Some of the expressed OMPs showed high binding activity with immunized rabbit antiserum.

Synergistic anti-tumor effect of ultrasound and hematoporphyrin on sarcoma180 cells with special reference to the changes of morphology and cytochrome oxidase activity of tumor cells.

This study is aimed at evaluating the inhibitory effects of the association of hematoporphyrin and ultrasound at variable intensities with a fixed frequency of 1.1MHz in tumor nodules. Specifically, the effects were studied both in solid and ascitic S180 tumors transplanted in mice by clinical, cytochemical and ultrastructural evaluation. The results indicated that the use of hematoporphyrin alone had no significant effect on destroying tumor cells. The ultrasound alone had little effect. Interestingly, the inhibition was much more effective when hematoporphyrin was combined with ultrasound. The inhibition was 3 times better than ultrasound alone and 8 times better than hematoporphyrin used alone. Our results also indicated that the changes on cell structure and cytochrome oxidation activity are important factors that could inhibit tumor cell growth and induce cell death. Apoptosis of tumor cells could be induced by hematoporphyrin. Our study investigated the killing mechanism on S180 tumor cells by using hematoporphyrin and low frequency ultrasound at cell, tissue and individual level.

Association of SNPs and haplotypes in GABAA receptor beta2 gene with schizophrenia.

Disturbances in GABAergic system have been observed in schizophrenics. In the present study, population association analysis was performed on 19 SNPs in the alpha(1), beta(2), gamma(2), epsilon and pi subunit genes of GABA(A) receptor. Five SNPs in GABRB2, namely B2I7G1584T, rs1816071, rs194072, rs252944 and rs187269, were found to be significantly associated, and their haplotypes in linkage disequilibrium, with schizophrenia. This represents the first report on any disease association of SNPs in the human GABA(A) receptor genes, and focuses attention on the GABAergic hypothesis of schizophrenia etiology.

Phylogeny Based on Whole Genome as inferred from Complete Information Set Analysis.

Previous molecular phylogeny algorithms mainly rely onmulti-sequence alignments of cautiously selected characteristic sequences,thus not directly appropriate for whole genome phylogeny where eventssuch as rearrangements make full-length alignments impossible. Weintroduce here the concept of Complete Information Set (CIS) and itsmeasurement implementation as evolution distance without reference tosizes. As method proof-test, the 16s rRNA sequences of 22 completelysequenced Bacteria and Archaea species are used to reconstruct aphylogenetic tree, which is generally consistent with the commonlyaccepted one. Based on whole genome, our further efforts yield a highlyrobust whole genome phylogenetic tree, supporting separate monophyleticcluster of species with similar phenotype as well as the early evolution ofthermophilic Bacteria and late diverging of Eukarya. The purpose of thiswork is not to contradict or confirm previous phylogeny standards butrather to bring a brand-new algorithm and tool to the phylogeny researchcommunity. The software to estimate the sequence distance and materialsused in this study are available upon request to corresponding author.

A new method for protein domain recognition.

A fuzzy cluster method is presented to recognize protein domains. This algorithm can identify domains globally. A protein structure set was used to test the algorithm. Among 219 proteins, 66.7% yielded results that agreed with the reference definitions, 30.6% showed minor differences, and only 2.7% (six proteins) showed major differences with the reference. The new method is more than 20 times fast than previous algorithms.

Modified penis lengthening surgery: review of 52 cases.

A modified surgical method for penis lengthening was, for the first time, set up in this laboratory. The procedure involves covering the dissected corpus cavernosum with either a scrotal flap or a skin graft after releasing the superficial ligament and even some deep suspensory ligament. The advantage of the scrotal flap is emphasized to cover the wound, and a V-Y suture was made to avoid the traction. The results, both in appearance and increased length, were satisfactory in 52 cases. Among the 52 patients, 39 suffered from congenital short penis and 13 from traumatic injuries. The significance and the blood supply of the penis are discussed.