RÉSUMÉ
Objective: Comparative analyses of wild-type Clostridioides difficile 630 (Cd630) strain and pathogenicity locus (PaLoc) knockout mutant (ΔPaLoc) by using RNA-seq technology. Analysis of differential expression of Cd630 wild-type strain and ΔPaLoc mutant strain and measurement of its cellular virulence changes. Lay the foundation for the construction of an toxin-attenuated vaccine strain against Clostridioides difficile. Methods: Analysis of Cd630 and ΔPaLoc mutant strains using high-throughput sequencing (RNA-seq). Clustering differentially expressed genes and screening differentially expressed genes by DESeq software. Further analysis of differential genes using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Finally, cytotoxicity assays of ΔPaLoc and Cd630 strains were performed in the African monkey kidney epithelial cell (Vero) and the human colonic cell (Caco-2) lines. Results: The transcriptome data showed that the ΔPaLoc mutant toxin genes tcdA and tcdB were not transcribed. Compared to the wild-type strain, CD630_36010, CD630_020910,CD630_02080 and cel genes upregulated 17.92,11.40,8.93 and 7.55 fold, respectively. Whereas the hom2 (high serine dehydrogenase), the CD630_15810 (spore-forming protein), CD630_23230 (zinc-binding dehydrogenase) and CD630_23240 (galactitol 1-phosphate 5-dehydrogenase) genes were down-regulated by 0.06, 0.075, 0.133 and 0.183 fold, respectively. The GO and KEGG enrichment analyses showed that the differentially transcribed genes in ΔPaLoc were enriched in the density-sensing system, ABC transport system, two-component system, phosphotransferase (PTS) system, and sugar metabolism pathway, as well as vancomycin resistance-related pathways. Cytotoxicity assays showed that the ΔPaLoc mutant strain lost its virulence to Vero and Caco-2 cells compared to the wild-type Cd630 strain. Conclusion: Transcriptional sequencing analysis of the Cd630 and ΔPaLoc mutant strains showed that the toxin genes were not transcribed. Those other differential genes could provide a reference for further studies on the physiological and biochemical properties of the ΔPaLoc mutant strain. Cytotoxicity assays confirmed that the ΔPaLoc mutant lost virulence to Vero and Caco-2 cells, thus laying the foundation for constructing an toxin-attenuated vaccine strain against C. difficile.
Sujet(s)
Toxines bactériennes , Clostridioides difficile , Protéines bactériennes/génétique , Protéines bactériennes/métabolisme , Toxines bactériennes/génétique , Toxines bactériennes/métabolisme , Cellules Caco-2 , Clostridioides , Clostridioides difficile/génétique , Humains , Oxidoreductases/génétique , Oxidoreductases/métabolisme , Transcriptome , Vaccins atténuésRÉSUMÉ
BACKGROUND: RNA-binding proteins have an important role in messenger RNA (mRNA) regulation during tumour development and carcinogenesis. In the present study, we examined the insulin-like growth factor 2 mRNA-binding proteins (IGF2BPs; hereafter refered to as IMPs) and Lin28 family expressions in epithelial ovarian carcinoma (EOC) patients and correlated their expression levels with the response to chemotherapy, hCTR1 expression and patient survival. METHODS: Patients clinical information, real-time RT-PCR, immunohistochemistry, western blot, Transwell migration invasion assays, and cytotoxicity assays were used. RESULTS: From 140 EOC patients, high expression of IMP3 or Lin28B was associated with poor survival, and women diagnosed at advanced stages with elevated IMP3 and Lin28B were at higher risk of developing chemoresistance. High IMP3 levels combined with high Lin28B levels significantly correlated with the poorest 5-year survival rates. Knockdown of IMP3 or Lin28B decreased cell proliferation, migration, and invasion, and increased the platinum sensitivity, but not taxol sensitivity, of ovarian cancer cells through increased expression of hCTR1, a copper transporter involved in platinum uptake. High expression of hCTR1 correlated with low expression of IMP3/Lin28B and better progression-free survival in advanced-stage EOC patients. CONCLUSION: Testing for a combination of elevated IMP3 and Lin28B levels could further facilitate the identification of a patient subgroup with the worst prognosis.
Sujet(s)
Résistance aux médicaments antinéoplasiques , Tumeurs épithéliales épidermoïdes et glandulaires/diagnostic , Tumeurs épithéliales épidermoïdes et glandulaires/mortalité , Tumeurs de l'ovaire/diagnostic , Tumeurs de l'ovaire/mortalité , Protéines de liaison à l'ARN/génétique , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Marqueurs biologiques tumoraux/génétique , Marqueurs biologiques tumoraux/métabolisme , Carcinome épithélial de l'ovaire , Lignée cellulaire tumorale , Résistance aux médicaments antinéoplasiques/génétique , Femelle , Régulation de l'expression des gènes tumoraux , Cellules HepG2 , Humains , Adulte d'âge moyen , Tumeurs épithéliales épidermoïdes et glandulaires/génétique , Tumeurs épithéliales épidermoïdes et glandulaires/anatomopathologie , Tumeurs de l'ovaire/génétique , Tumeurs de l'ovaire/anatomopathologie , Pronostic , Protéines de liaison à l'ARN/métabolisme , Taux de survie , Régulation positive/génétiqueRÉSUMÉ
OBJECTIVES: To investigate, using three-dimensional power Doppler ultrasound (3D-PDU), alterations in cervical intratumoral vascularization during and after radiotherapy. METHODS: Between 2004 and 2009 we enrolled into the study 37 patients with FIGO Stages IB1-IIB cervical carcinoma who were undergoing radiotherapy. Serial 3D-PDU scans were performed during treatment, providing ultrasonographic measurement of tumor size, vascularization index, flow index and vascularization flow index, as well as monthly for 3 months post-treatment and tri-monthly thereafter, until vascularity was undetectable on two consecutive occasions. Physical examination, cervical cytology and serum marker evaluation were performed every 3-6 months for the first 5 years following treatment. Patients evaluated after a 2-year tumor-free interval and those with clinically assessed positive findings at follow-up underwent 3D-PDU to detect possible local disease. RESULTS: A total of 329 3D-PDU scans were performed in the 37 women. Cervical tumors and intratumoral vascularization disappeared within 3 months following radiotherapy, except in one patient with persistent disease. Nine patients had disease relapse, in four of whom the recurrence was local. In three of these four, there was recurrence of tumor and vascularization after a complete response. At follow-up, 3D-PDU detected local disease with 75.0% sensitivity and 98.5% specificity, while serum markers detected local disease among 34 patients with squamous cell carcinoma with 20.0% sensitivity and 77.3% specificity. CONCLUSIONS: Compared with serum markers in cervical squamous cell carcinoma, 3D-PDU has higher sensitivity and specificity for detecting local recurrence or persistence in cervical carcinoma. Thus, 3D-PDU combined with clinical assessment may be a new and safe method for monitoring radiotherapy treatment response and detecting local recurrence.
Sujet(s)
Carcinome épidermoïde/anatomopathologie , Col de l'utérus/anatomopathologie , Imagerie tridimensionnelle , Récidive tumorale locale/anatomopathologie , Examen physique , Tumeurs du col de l'utérus/anatomopathologie , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Marqueurs biologiques/sang , Vitesse du flux sanguin , Carcinome épidermoïde/imagerie diagnostique , Carcinome épidermoïde/radiothérapie , Col de l'utérus/vascularisation , Femelle , Études de suivi , Humains , Adulte d'âge moyen , Néovascularisation pathologique , Études prospectives , Sensibilité et spécificité , Résultat thérapeutique , Charge tumorale , Échographie-doppler couleur , Tumeurs du col de l'utérus/imagerie diagnostique , Tumeurs du col de l'utérus/radiothérapie , Vagin/imagerie diagnostiqueRÉSUMÉ
The rumen content of four Yunnan Yellow Cattle (Bos taurs) were collected to determine the bacteria diversity by using 16S rRNA gene sequence analysis. A total of 129 sequences were examined and the sequences were referred as 107 OTU (Operational Taxonomy Unit) according to the similarity level of 97% in gene sequence. Similarity analysis revealed that Yunnan Yellow Cattle had 12 sequences (10 OTU) shared 97% or greater similarity with cultured rumen bacteria Butyrivibrio fibrisolvens, Succiniclasticum ruminis, Ruminococcus bromii, Clostridium proteoclasticum, Ruminococcus flavefaciens, Pseudobutyrivibrio ruminis, Jeotgalicoccus psychrophilus, and Prevotella ruminicola, which accounting for 9.3% of the total clones (9.2% of the total OTU). The further 12 sequences (9 OTU) shared 90-97% similarity with cultured bacteria Clostridium aminobutyricum, butyrate-producing bacterium, Schwartzia succinivorans, Prevotella ruminicola, Eubacterium ruminantium, Ruminococcus albus, and Clostridium termitidis, also accounting for 9.3% of the total sequences (8.3% of the total OTU). The remaining 105 sequences (90 OTU) shared less than 90% similarity with cultured bacteria, accounting for 81.4% of the total sequences (82.5% of the total OTU). According to the phylogenetic analysis, all sequences were phylogenetically placed within phyla of low G+C subdivision (accounting for 72.1 and 72.5% of the total clones and OTU, respectively) and CFB subdivision (Cytophaga-Flexibacter-Bacteroides; accounting for 27.9 and 27.5% of the total clones and OTU, respectively). Among the examined clones, rare bacteria Jeotgalicoccus psychrophilus was detected in the rumen of cattle.
Sujet(s)
Bactéries/génétique , Bovins/microbiologie , Variation génétique , Phylogenèse , Rumen/microbiologie , Animaux , Bactéries/classification , Séquence nucléotidique , Chine , Clonage moléculaire , Analyse de regroupements , Données de séquences moléculaires , ARN ribosomique 16S/génétique , Analyse de séquence d'ADN , Similitude de séquencesRÉSUMÉ
Kv2.1 channels exhibit a U-shaped voltage-dependence of inactivation that is thought to represent preferential inactivation from preopen closed states. However, the molecular mechanisms underlying so-called U-type inactivation are unknown. We have performed a cysteine scan of the S3-S4 and S5-P-loop linkers and found sites that are important for U-type inactivation. In the S5-P-loop linker, U-type inactivation was preserved in all mutant channels except E352C. This mutation, but not E352Q, abolished closed-state inactivation while preserving open-state inactivation, resulting in a loss of the U-shaped voltage profile. The reducing agent DTT, as well as the C232V mutation in S2, restored U-type inactivation to the E352C mutant, which suggests that residues 352C and C232 may interact to prevent U-type inactivation. The R289C mutation, in the S3-S4 linker, also reduced U-type inactivation. In this case, DTT had little effect but application of MTSET restored wild-type-like U-type inactivation behavior, suggestive of the importance of charge at this site. Kinetic modeling suggests that the E352C and R289C inactivation phenotypes largely resulted from reductions in the rate constants for transitions from closed to inactivated states. The data indicate that specific residues within the S3-S4 and S5-P-loop linkers may play important roles in Kv2.1 U-type inactivation.
Sujet(s)
Canaux potassiques Shab/composition chimique , Canaux potassiques Shab/métabolisme , Animaux , Disulfures/composition chimique , Cinétique , Modèles biologiques , Mutation , Rats , Canaux potassiques Shab/génétiqueRÉSUMÉ
Suicidal ideation is life-threatening and is prevalent among people living with HIV (PLWH). A dearth of such studies was conducted in China. This study investigated the prevalence of suicidal ideation and associated factors among PLWH who were former blood and/or plasma donors (FBPD) in a rural county in central China. Prospective respondents were randomly selected from a local registry; 176 PLWH who were FBPD joined the study. With informed consent, these PLWH and their spouse were interviewed separately and anonymously. Respectively, 34 and 8% of the sampled (index) PLWH self-reported having suicidal ideation and making a suicidal attempt in the last year. The multivariate analyses results showed that the index PLWH's Physical Function subscale score of the Medical Outcomes Study HIV Health Survey scale (Odds ratio (OR)=6.67, 95% CI=1.69-26.27,
Sujet(s)
Donneurs de sang/psychologie , Infections à VIH/psychologie , Idéation suicidaire , Adolescent , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Donneurs de sang/statistiques et données numériques , Chine/épidémiologie , Femelle , Infections à VIH/épidémiologie , Séropositivité VIH/épidémiologie , Séropositivité VIH/psychologie , Enquêtes de santé , Humains , Mâle , Adulte d'âge moyen , Analyse multifactorielle , Facteurs de risque , Santé en zone rurale , Facteurs socioéconomiques , Jeune adulteRÉSUMÉ
A random sample of 271 people living with HIV (PLWH) who were former plasma/blood donors and a convenience sample of 67 HIV negative villagers were anonymously interviewed. Compared with the non-PLWH, PLWH reported higher prevalence of symptoms of depression (adjusted OR = 2.53, p=0.001), anxiety (adjusted OR = 1.85, p=0.04), and stress (adjusted OR = 1.77, p=0.06). Of the PLWH respondents, 81.7% received Highly Active Antiretroviral Therapy (HAART); 32.1% of whom reported some side effects. Respectively 13.7%, 37.4%, and 38.4% PLWH perceived discrimination from their family members, relatives/friends, and neighbors. Absence of HAART, poor physical function, perceived discrimination from relatives and friends, and low level of resilience were associated with depression (stepwise regression; beta = - 0.28-0.17, R-square = 0.22), anxiety and stress (R-square = 0.32 and 0.16, respectively). The majority of respondents (70.1%) desired group intervention as a means for providing psychological support services. Relevant programs should both remove risk factors (e.g., absence of medical treatment, HIV-related discrimination) and promote protective factors (e.g., resilience). Support group is one of the potentially useful approaches to provide psychological support services.
Sujet(s)
Thérapie antirétrovirale hautement active/psychologie , Donneurs de sang , Trouble dépressif/psychologie , Infections à VIH/psychologie , Stéréotypes , Adolescent , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Études cas-témoins , Chine/épidémiologie , Trouble dépressif/traitement médicamenteux , Trouble dépressif/épidémiologie , Trouble dépressif/étiologie , Femelle , Infections à VIH/complications , Infections à VIH/traitement médicamenteux , Infections à VIH/épidémiologie , Humains , Mâle , Adulte d'âge moyen , Plasma sanguin , Prévalence , Facteurs de risque , Santé en zone rurale , Stress psychologique , Jeune adulteRÉSUMÉ
This study was aimed to identify the expression and the correlation of insulin-like growth factor-1 (IGF-1) system and their prognostic impacts in cervical cancer. Seventy-two patients with early-stage cervical cancer were eligible. We obtained the serum levels of total IGF-1 and IGF binding protein-3 (IGFBP-3) by enzyme-linked immunosorbent assay and the expression of IGF-1 receptor (IGF-1R) in cancerous tissue by immuno-fluorescent (IF) stains. The 5-year recurrence-free and overall survival rates were significantly lower (P=0.003 and P=0.01, respectively) among patients with high-grade expression of tissue IGF-1R, compared with those with low-grade expression. After adjustment for other factors, preoperative serum total IGF-1 or IGFBP-3 levels failed to predict cancer death and recurrence. High-grade expression of IGF-1R and elevated preoperative squamous cell carcinoma antigen level were independent predictors of both death and recurrence, and combination of both factors could further help identify the subgroup of patients at higher death risk. The IF staining indicates the colocalisation of IGF-1 and IGF-1R in the cancerous tissues, whereas the IGF-1R expression is not correlated with circulating levels of IGF-1 or IGFBP-3. In early-stage cervical cancer, IGF-1 system may have a paracrine or autocrine function and the adverse impacts on prognosis by IGF-1R overexpression are implicated.
Sujet(s)
Facteur de croissance IGF-I/physiologie , Tumeurs du col de l'utérus/physiopathologie , Femelle , Technique d'immunofluorescence , Humains , Protéine-3 de liaison aux IGF/sang , Analyse multifactorielle , Pronostic , Récepteur IGF de type 1/métabolisme , Transduction du signal , Taux de survie , Tumeurs du col de l'utérus/anatomopathologieRÉSUMÉ
We examined the potential contribution of a voltage-gated proton conductance (gH+) to acid extrusion from cultured postnatal rat hippocampal neurons. In neurons loaded with Ca2+- and/or pH-sensitive fluorophores, transient exposures to 25-139.5 mM external K+ (K+o) or 20 microM veratridine in the presence of 2 mM Ca2+o (extracellular pH (pHo) constant at 7.35) caused reversible increases and decreases in intracellular free calcium concentration ([Ca2+]i) and intracellular pH (pHi), respectively. In contrast, under external Ca2+-free conditions, the same stimuli failed to affect [Ca2+]i but caused an increase in pHi, the magnitude of which was related to the [K+]o applied and the change in membrane potential. Consistent with the properties of gH+s in other cell types, the magnitude of the rise in pHi observed in the absence of external Ca2+ was not affected by the removal of external Na+ but was sensitive to external Zn2+ and temperature and was dependent on the measured transmembrane pH gradient (DeltapHmemb). Increasing DeltapH(memb) by pretreatment with carbonylcyanide-p-trifluoromethoxyphenylhydrazone augmented both the high-[K+]o-evoked rise in pHi and the Zn2+-sensitive component of the rise in pHi, suggestive of increased acid extrusion via a gH+. The inhibitory effect of Zn2+ at a given DeltapHmemb was further enhanced by increasing pHo from 7.35-7.8, consistent with a pHo-dependent inhibition of the putative gH+ by Zn2+. Under conditions designed to isolate H+ currents, a voltage-dependent outward current was recorded from whole-cell patch-clamped neurons. Although the outward current appeared to show some selectivity for protons, it was not sensitive to Zn2+ or temperature and the H+-selective component could not be separated from a larger conductance of unknown selectivity. Nonetheless, taken together, the results suggest that a Zn2+-sensitive proton conductive pathway is present in rat hippocampal neurons and contributes to H+ efflux under depolarizing conditions.
Sujet(s)
Acides/métabolisme , Hippocampe/cytologie , Ouverture et fermeture des portes des canaux ioniques/physiologie , Neurones/physiologie , Protons , Animaux , Animaux nouveau-nés , Calcium/pharmacologie , ([4-(Trifluorométhoxy)phényl]hydrazono)malononitrile/pharmacologie , Cellules cultivées , Interactions médicamenteuses , Éosine B , Fluorescéines/pharmacologie , Ouverture et fermeture des portes des canaux ioniques/effets des médicaments et des substances chimiques , Ouverture et fermeture des portes des canaux ioniques/effets des radiations , Ionophores/pharmacologie , Spectrométrie de masse , Potentiels de membrane/effets des médicaments et des substances chimiques , Potentiels de membrane/physiologie , Potentiels de membrane/effets des radiations , Neurones/effets des médicaments et des substances chimiques , Techniques de patch-clamp , Potassium/pharmacologie , Rats , Rat Wistar , Vératridine/pharmacologieRÉSUMÉ
Evolutionary conservation derived from a multiple sequence alignment is a powerful indicator of the functional significance of a residue, and it can help to predict active sites, ligand-binding sites, and protein interaction interfaces. The results of the existing algorithms in identifying the residue's conservation strongly depend on the sequence alignment, making the results highly variable. Here, by introducing the amino acid similarity matrix, we propose a novel gap-treating approach by combining the evolutionary information and von Neumann entropies to compute the residue conservation scores. It is indicated through a series of tested results that the new approach is quite encouraging and promising and may become a useful tool in complementing the existing methods.
Sujet(s)
Biologie informatique/méthodes , Séquence conservée , Entropie , Évolution moléculaire , Protéines/composition chimique , Analyse de séquence de protéine/méthodes , Algorithmes , Acides aminés/composition chimique , Sites de fixation , Simulation numérique , Bases de données de protéines , Ligands , Valeur prédictive des tests , Protéines/génétique , Sensibilité et spécificité , Alignement de séquencesRÉSUMÉ
Squamous cell carcinoma antigen (SCCA) is a tumor marker for patients with squamous cell carcinoma of uterine cervix, lung, and esophagus. It was encoded by two highly homologous genes, SCCA1 and SCCA2. However, the relevance of SCCA genes to squamous cell carcinogenesis and patient outcome remains far from clear. In this study, by using laser microdissection and real-time quantitative polymerase chain reaction procedures, the messenger RNA (mRNA) expression of the SCCA1 and SCCA2 genes in normal, dysplastic, and malignant squamous epithelia from uterine cervical tissues were analyzed and correlated with outcome of cancer patients. We found that the SCCA2/A1 mRNA ratios were progressively increased from normal, dysplastic, to cancer cells, and the mean ratio was significantly higher in cancer tissues than that in normal epithelium (P= 0.02). The SCCA2/A1 mRNA ratios were not significantly associated with types of human papillomavirus infection (P > 0.05). High SCCA2/SCCA1 mRNA ratios (ratio >1) were an independent predictor of disease recurrence (relative risk: 3.58; P= 0.003). Of the 38 patients with cervical cancer, 12 patients with high SCCA2/SCCA1 mRNA ratios had a significant lower 2-year disease-free survival of only 50%, while it was 92% in those with low SCCA2/SCCA1 mRNA ratios (P < 0.001). In conclusion, our study indicated that the ratios of SCCA2 to SCCA1 RNA were increased during the process of cervical carcinogenesis, and patients with elevated SCCA2/A1 ratio carried a higher risk for recurrence in early-stage uterine cervical cancer.
Sujet(s)
Antigènes néoplasiques/biosynthèse , Marqueurs biologiques tumoraux/biosynthèse , Serpines/biosynthèse , Tumeurs du col de l'utérus/immunologie , Antigènes néoplasiques/génétique , Marqueurs biologiques tumoraux/génétique , Femelle , Études de suivi , Humains , Adulte d'âge moyen , Pronostic , ARN messager/biosynthèse , ARN messager/génétique , RT-PCR , Serpines/génétique , Tumeurs du col de l'utérus/génétiqueRÉSUMÉ
As more and more genomes have been discovered in recent years, there is an urgent need to develop a reliable method to predict the subcellular localization for the explosion of newly found proteins. However, many well-known prediction methods based on amino acid composition have problems utilizing the sequence-order information. Here, based on the concept of Chou's pseudo amino acid composition (PseAA), a new feature extraction method, the multi-scale energy (MSE) approach, is introduced to incorporate the sequence-order information. First, a protein sequence was mapped to a digital signal using the amino acid index. Then, by wavelet transform, the mapped signal was broken down into several scales in which the energy factors were calculated and further formed into an MSE feature vector. Following this, combining this MSE feature vector with amino acid composition (AA), we constructed a series of MSEPseAA feature vectors to represent the protein subcellular localization sequences. Finally, according to a new kind of normalization approach, the MSEPseAA feature vectors were normalized to form the improved MSEPseAA vectors, named as IEPseAA. Using the technique of IEPseAA, C-support vector machine (C-SVM) and three multi-class SVMs strategies, quite promising results were obtained, indicating that MSE is quite effective in reflecting the sequence-order effects and might become a useful tool for predicting the other attributes of proteins as well.
Sujet(s)
Algorithmes , Acides aminés/analyse , Simulation numérique , Structure quaternaire des protéines , Protéines/composition chimique , Protéines/classification , Bases de données factuelles , Valeur prédictive des tests , Pliage des protéinesRÉSUMÉ
Molecular cytogenetic analysis identified a new type of dicentric chromosome involving different breakpoints at 18q in a female fetus. The chromosome anomaly was designated as an asymmetrical pseudoisodicentric chromosome 18, 46,XX,psu dic(18)(pter-->q11.2::q21.3-->pter)mat. A series of BAC clones for 18q11.2 and q21.3 regions were used to identify one breakpoint within the region q11.2 between 19.8 and 21.6 Mb from the telomere of 18p and another breakpoint within q21.3 between 55.4 and 56.9 Mb from the telomere of 18p by FISH analysis. Real-time quantitative PCR and microsatellite analysis further verified that the dicentric chromosome was maternal in origin and resulted from a break-reunion between sister chromatids of a single maternal chromosome. We propose that a loop-type configuration of sister chromatids took place and that the break-reunion occurred at cross sites of the loop to form an asymmetrical isodicentric chromosome during either mitosis or meiosis. In this case, the asymmetrical pseudoisodicentric resulted in an 18pter--> q11.2 duplication and an 18q21.3-->qter deletion, which could have led to certain dysmorphic features of 18q- syndrome in this fetus.
Sujet(s)
Aberrations des chromosomes , Chromosomes humains de la paire 18/génétique , Adulte , Chromosomes artificiels de bactérie , Clones cellulaires , Femelle , Foetus/malformations , Humains , Hybridation fluorescente in situ , Caryotypage , Répétitions microsatellitesRÉSUMÉ
A minilibrary was constructed from DOP-PCR products using microdissected Y-chromosomes of Indian muntjac as DNA templates. Two microclones designated as IM-Y4-52 and IM-Y5-7 were obtained from negative screening of all three cervid satellite DNAs (satellites I, II, and IV). These two microclones were 295 and 382 bp in size, respectively, and shared approximately 70% sequence homology. Southern blot analysis showed that the IM-Y4-52 clone was repetitive in nature with an approximately 0.32-kb register in HaeIII digest. Sequence comparison revealed no similarities to DNA sequences deposited in the GenBank database, suggesting that the microclone sequences were from a novel satellite DNA family designated as cervid satellite V. A subclone of an Indian muntjac BAC clone which screened positive for IM-Y4-52 had a 3,325-bp insert containing six intact monomers, four deleted monomers, and two partial monomers. The consensus sequence of the monomer was 328 bp in length and shared more than 80% sequence homology with every intact monomer. A zoo blot study using IM-Y4-52 as a probe showed that the strong hybridization with EcoRI digested male genomic DNA of Indian muntjac, Formosan muntjac, Chinese muntjac, sambar deer, and Chinese water deer. Female genomic DNA of Indian muntjac, Chinese water deer, and Formosan muntjac also showed positive hybridization patterns. Satellite V was found to specifically localize to the Y heterochromatin region of the muntjacs, sambar deer, and Chinese water deer and to chromosome 3 of Indian muntjac and the X-chromosome of Chinese water deer.
Sujet(s)
ADN satellite , Muntiacus/génétique , Chromosome Y/génétique , Animaux , Séquence nucléotidique , Évolution biologique , Chromosomes/ultrastructure , Femelle , Hybridation fluorescente in situ , Caryotypage , Mâle , Microdissection , Modèles génétiques , Séquences répétées d'acides nucléiques , Similitude de séquences d'acides nucléiquesRÉSUMÉ
OBJECTIVES: To characterize intratumoral vascularization in early-stage cervical cancer by three-dimensional (3D) power Doppler ultrasound. METHODS: One hundred and forty-one patients with carcinoma of the uterine cervix and 30 normal controls were studied by transvaginal 3D power Doppler ultrasound. The tumor volume of the cervical cancer was determined. The blood flow within the tumor or normal cervix was measured and expressed as the vascularization index (VI), flow index (FI) and vascularization flow index (VFI). RESULTS: Of the 141 patients with cervical cancer, 44 patients had undergone prior cervical conization. Eighty-seven patients had measurable cervical tumors, of whom five had had prior conization. Abundant intratumoral power Doppler signals could be detected, and the VI, FI and VFI were significantly elevated in cervical cancer patients compared with women with a normal cervix and patients in whom no cervical tumor could be detected (P < 0.05, one-way ANOVA). We observed four types of intratumoral vascularity patterns, which did not significantly differ in VI, FI and VFI: localized, peripheral, scattered and single-vessel types. Cervical tumor volume was positively correlated with FI (linear regression, r = 0.373, P = 0.001), but not with VI or VFI. CONCLUSIONS: 3D power Doppler ultrasound provides a useful tool to investigate intratumoral vascularization and volume of cervical cancer.
Sujet(s)
Échographie-doppler/méthodes , Tumeurs du col de l'utérus/imagerie diagnostique , Adulte , Sujet âgé , Analyse de variance , Diagnostic précoce , Femelle , Humains , Imagerie tridimensionnelle/méthodes , Adulte d'âge moyen , Biais de l'observateur , Tumeurs du col de l'utérus/vascularisationRÉSUMÉ
Haglund's deformity, or "pump bump" is a common cause of posterior heel pain, characterized clinically by a painful soft tissue swelling at the level of the Achilles tendon insertion. We reviewed 30 heels in 19 patients with failure of conservative treatment. Surgical management consisted of excision of the posterior calcaneal tuberosity and bursectomy through a medial longitudinal incision. The average follow-up period was 6 years (range, 3-10 years). Only 3 heels (10%) in 2 patients had persistent pain. Twenty-seven heels (90%) were cured after operation. However, 25 heels (83%) had residual pain for a half to two years after operation and became free of symptoms thereafter. We conclude surgical treatment of Haglund's deformity produces a predictably good result (90%) but requires a long time about a half to two years for full recovery.
Sujet(s)
Anomalies morphologiques du pied/chirurgie , Talon , Adolescent , Adulte , Femelle , Humains , Mâle , Adulte d'âge moyen , Douleur/étiologieRÉSUMÉ
We have reviewed a single surgeon's experience with isolated talonavicular fusion in 16 patients with talonavicular arthritis. Fixation was either by staples or screws. Fifteen solid unions were achieved in the 16 patients who were followed (mean: 51 months). The average Ankle-Hindfoot Scale improved from 77.2 preoperatively to 92.9 postoperatively (p < 0.001). Subjectively, 15 patients were satisfied and one patient dissatisfied with the results. Further osteoarthritis in the adjacent joints was noted in five patients. We concluded that isolated talonavicular fusion is an effective method of treatment of talonavicular arthritis regarding pain relief and functional improvement. Though osteoarthritis was found in some adjacent joints postoperatively, the results were still satisfactory.
Sujet(s)
Arthrite/chirurgie , Arthrodèse/méthodes , Talus/chirurgie , Os du tarse/chirurgie , Adolescent , Adulte , Arthrite/imagerie diagnostique , Femelle , Études de suivi , Humains , Mâle , Adulte d'âge moyen , Arthrose/chirurgie , Satisfaction des patients , RadiographieRÉSUMÉ
Concomitant ipsilateral femoral shaft and neck fractures are difficult to treat. The standard protocol in our hospital is plates for diaphyseal fractures and lag screws or dynamic hip screws (DHS) fixations for the femoral neck fractures. We treated 21 patients with these complex fractures between 1988 and 1998 with this protocol. All patients were injured from high-energy trauma. There were 15 males and 6 females with an average age of 44 years. The average follow-up period is 50 months. All except one patient united well. During the follow-up period, neither non-union of the femoral neck fracture nor osteonecrosis of the femoral head was noted. There was also no malunion. There were twenty good results and one poor functional result. We conclude that plate on the shaft and sliding hip screws or separate screws in the hip are a reliable method for concomitant ipsilateral femoral neck and shaft fractures.