Obstetrical Outcomes of Amniocentesis or Chorionic Villus Sampling in Dichorionic Twin Pregnancies

BACKGROUND: Under certain situations, women with twin pregnancies may be counseled to undergo invasive prenatal diagnostic testing. Chorionic villus sampling and amniocentesis are the two generally performed invasive prenatal diagnostic tests. Studies comparing procedure-related fetal loss between first-trimester chorionic villus sampling and second-trimester amniocentesis in twin pregnancies are limited. This study aimed to evaluate the procedure-related fetal loss and the obstetrical outcomes of these two procedures, chorionic villus sampling and amniocentesis in twin pregnancies. METHODS: The data from dichorionic-diamniotic twin pregnancies on which first-trimester chorionic villus sampling (n = 54) or second-trimester amniocentesis (n = 170) was performed between December 2006 and January 2017 in a single center were retrospectively analyzed. The procedure-related fetal loss was classified as loss of one or all fetuses within 4 weeks of procedure, and overall fetal loss was classified as loss of one or all fetuses during the gestation. The groups were compared with respect to the procedure-related and obstetrical outcomes. RESULTS: The difference in proportion of procedure-related fetal loss rate (1.9% for chorionic villus sampling vs. 1.8% for amniocentesis; P = 1.000) and the overall fetal loss rate (7.4% for chorionic villus sampling vs. 4.7% for amniocentesis; P = 0.489) between the two groups was not significant. The mean gestational ages at delivery were not statistically significant. CONCLUSION: Both the overall fetal loss rate and the procedure-related fetal loss rate of chorionic villus sampling and amniocentesis in dichorionic twin pregnancies had no statistical significance. Both procedures can be safely used individually.

Higroma quístico retronucal como marcador de anomalías cromosómicas en el primer trimestre de la gestación / Retronuchal cystic hygroma in the first trimester of pregnancy as a marker of chromosomal abnormalities

Background: Nuchal cystic hygroma is the most frequently identified marker of chromosomal anomalies during first trimester screening. Objective: To determine the association of the nuchal cystic hygroma with chromosomal anomalies diagnosed with karyotyping done between the first and second trimesters of pregnancy. Design: Retrospective study. Setting: Instituto Latinoamericano de Salud Reproductiva (ILSAR), Lima, Peru. Patients. Fetuses with nuchal cystic hygroma. Methods: The data were obtained from the ILSAR database between August 2007 and May 2018, the cases diagnosed by ultrasound from week 11 to 13.6. Nuchal cystic hygroma was defined as the presence of septated liquid content in the nuchal axial section with a thickness above the 95th percentile value for increased nuchal translucency value for the crown-rump length. The karyotype was obtained between the first and second trimesters from material collected by chorionic villus sampling (BVS) or amniocentesis (AMC). Main outcome measures: Karyotyping results were compared between cases with cystic hygroma alone and cases with cystic hygroma in addition to another marker. Results: Out of 459 invasive procedures performed in fetuses with high risk for chromosomal anomalies based on the Fetal test database of Spain, there were 162 cases of chromosomal anomalies (35.3%), and 104 cases of nuchal cystic hygroma (22.7%). Nuchal cystic hygroma was associated with a higher frequency of chromosomal abnormalities, compared to fetuses without cystic hygroma (52.9% vs. 30.1%; p<0.001). Out of 61 cases of hygroma alone, 42.3% had chromosomal anomalies, and when the hygroma was associated with other markers (fetal hydrops, abnormal ductus venosus, heart disease), 65.1% had chromosomal abnormalities. There was a statistically significant difference (p=0.003) for the presence of monosomy X between the group with cystic hygroma alone and the group with hygroma and fetal hydrops. There was no difference in hygroma thickness between the groups with and without chromosomal abnormalities. Conclusions: Nuchal cystic hygroma is a risk marker with high predictive value for chromosomal abnormalities, and its identification during prenatal screening may be considered an indication to a diagnostic test. When cystic hygroma is associated to flow abnormalities of the ductus venosus or fetal hydrops, chromosomal abnormalities significantly increase. The hygroma associated with hydrops was primarily linked to monosomy X, while the hygroma associated with abnormal flow velocity waveforms of the ductus venosus was linked to trisomy 21.

Antecedentes. El higroma quístico retronucal es el marcador de anomalías cromosómicas identificado con mayor frecuencia en el tamizaje del primer trimestre. Objetivo. Evaluar la asociación del higroma quístico retronucal y anomalías cromosómicas diagnosticadas con el cariotipo, entre el primer y segundo trimestre del embarazo. Diseño. Estudio retrospectivo. Institución. Instituto Latinoamericano de Salud Reproductiva (ILSAR), Lima, Perú. Pacientes. Fetos con higroma quístico retronucal. Método. Estudio de fetos con higroma quístico retronucal, obtenidos de la base de datos de ILSAR, entre agosto del 2007 y mayo del 2018, diagnosticados por ecografía entre las 11 y 13,6 semanas. El higroma quístico retronucal se definió como la presencia de contenido líquido tabicado en el corte axial retronucal con un grosor mayor al percentil 95 del valor de translucencia nucal aumentada para la longitud corona-nalga. Se obtuvo el cariotipo entre el primer y segundo trimestre en material obtenido por biopsia de vellosidades coriales (BVC) o amniocentesis (AMC). Principales medidas de resultados. Los resultados del cariotipo fueron comparados entre los casos de higroma quístico solo y los casos que tuvieron higroma y adicionalmente otro marcador. Resultados. De un total de 459 procedimientos invasivos realizados en fetos con alto riesgo para anomalías cromosómicas en base al Fetal test de España, hubieron 162 casos de anomalías cromosómicas (35,3%) y se identificó 104 casos de higroma quístico retronucal (22s7%). El hallazgo de higroma quístico retronucal se asoció con mayor presencia de anomalías cromosómicas, comparado con los fetos sin higroma quístico (52,9% vs. 30,1%; p<0,001). De 61 casos de higroma solo, 42,3% tenían anomalía cromosómica, y cuando el higroma estaba asociado a otros marcadores (hidrops fetal, ductus venoso anormal, cardiopatía, ausencia de hueso nasal), hubo 65,1% de anomalías cromosómicas. Hubo diferencia estadística significativa (p=0,003) para la presencia de monosomía X, entre el grupo con higroma solo y el de higroma + hidrops fetal. No hubo diferencia en el grosor del higroma entre el grupo con y sin anomalía cromosómica. Conclusiones. El higroma quístico retronucal es un marcador de riesgo con alto valor predictivo para anomalías cromosómicas. Su identificación en el tamizaje prenatal podría ser indicación para recomendar una prueba diagnóstica. Cuando se asocia a anormalidad del flujo del ductus venoso o hidrops fetal, aumentan significativamente las anomalías cromosómicas. El higroma asociado con hidrops se vinculó mayoritariamente a la monosomía X, mientras que el higroma asociado con onda de velocidad de flujo-OVF de ductus venoso anormal a la trisomía 21.

Amniocentesis precoz y biopsia de vellosidad corial. Pérdidas fetales y anomalías congénitas en un grupo de gestantes brasileñas / Early amniocentesis and chorionic villus biopsy. Fetal losses and congenital anomalies in a group of Brazilian pregnant women

Resumen En el Instituto de Medicina Fetal y Genética Humana de São Paulo se ofrecen, a las gestantes que tienen un riesgo aumentado para anomalías cromosómicas, diferentes técnicas, entre ellas, la Biopsia de Vellosidad Corial Transabdominal (BVCTA) y la Amniocentesis Precoz (AP). El objetivo del presente estudio es realizar una comparación de la frecuencia de pérdidas fetales y anomalías congénitas presentadas en cada uno de los procedimientos, ambos realizados por los mismos operadores, en la misma edad gestacional (12-14 6/7 semanas) y bajo un abordaje transabdominal. Fueron analizadas retrospectivamente 432 AP y 418 BVCTA. Todos los procedimientos de colecta fueron monitorizados por ultrasonografía. La frecuencia de pérdidas fetales espontáneas fue del 4,9 % en AP y de 5,3 % en BVCTA, una diferencia no significativa. No se encontraron diferencias significativas entre los dos procedimientos al comparar las frecuencias de pérdidas en cada semana de gestación. Sangrado y pérdida de líquido amniótico fueron más frecuentes en AP que en la BVCTA. Esa diferencia fue significativa en el caso de la pérdida de líquido amniótico. En algunos casos este hallazgo se relacionó con pérdida fetal. La incidencia de prematuridad y bajo peso al nacimiento no difirió significativamente entre los dos procedimientos. La mayor frecuencia de problemas respiratorios registrada en AP no fue significativa en comparación con BVCTA. No se observó diferencia significativa en la incidencia de anomalías músculo-esqueléticas. La amniocentesis después de catorce semanas presenta un bajo riesgo de pérdida fetal o anomalías congénitas. La BVCTA, debe ser realizada alrededor de la semana doce de gestación.

Abstract In the Institute of Fetal Medicine and Human Genetics of São Paulo are offered, pregnant women have an increased risk for chromosomal abnormalities, different techniques, among them, Transabdominal Corial Vellosity Biopsy (BVCTA) and Precocious Amniocentesis (AP). The objective of this study is to compare the frequency of Hepatitis and congenital anomalies presented in all procedures, both performed by operators, in the same gestational age (12-14 6/7 weeks) and under a transabdominal approach. 432 AP and 418 BVCTA were analyzed retrospectively. All collection procedures were monitored by ultrasonography. The spontaneous fetal frequency was 4.9% in AP and 5.3% in BVCTA, a non-significant difference. There is no difference in results compared to gestation times. Bleeding and loss of amniotic fluid were more frequent in AP than in BVCTA. That difference was significant in the case of the loss of amniotic fluid. In some cases, this finding was related to fetal loss. The incidence of prematurity and birth weight without difference between the two procedures. The highest frequency of respiratory problems recorded in AP was not significant compared to BVCTA. There is no significant difference in the incidence of musculoskeletal abnormalities. Amniocentesis after 14 weeks presents a low risk of fetal loss or congenital anomalies. The BVCTA should be close to the twelfth week of gestation.

Expression of TET and 5-HmC in Trophoblast Villi of Women with Normal Pregnancy and with Early Pregnancy Loss / 华中科技大学学报（医学）（英德文版）

Increasing evidence suggests that epigenetic dysfunction may influence the stability of normal pregnancy.The ten-eleven translocation (TET) family and 5-hydroxymethylcytosine (5-hmC) were found to be linked with epigenetic reprogramming.The present study aimed to examine the expression of the TET family and 5-hmC in the villi of human embryos and compared their expression between normal pregnancy and early pregnancy loss (EPL).Embryonic villi were collected from normal pregnant women (control) experiencing medical abortion and from EPL patients at gestation ages of 6,7 and 8 weeks.The mRNAs of TET family were analysed using quantitative polymerase chain reaction (qPCR),and TET proteins using Western blotting and immunohistochemical analysis.The MethylFlashTM Kit was used to quantify the absolute amount of 5-methylcytosine (5-mC) and 5-hmC.Our results showed that the expression of the TETs and 5-hmC in the normal villus decreased with increasing gestational age.Immunohistochemistry revealed that the TET proteins were expressed in the cytoplasm of trophoblasts and their expression was the highest in the 6-week tissue samples,which was consistent with the qPCR and Western blot results.The expression of TET1,TET2,and TET3 was lower in the villi in EPL group than in normal pregnancy group (P＜0.05 for all).It was concluded that the TET family and 5-hmC are critical in epigenetic reprogramming of human embryo.The findings also suggest that a deficiency of TETs in the villus might be associated with human EPL.

Expression of TET and 5-HmC in Trophoblast Villi of Women with Normal Pregnancy and with Early Pregnancy Loss / 华中科技大学学报（医学）（英德文版）

Increasing evidence suggests that epigenetic dysfunction may influence the stability of normal pregnancy.The ten-eleven translocation (TET) family and 5-hydroxymethylcytosine (5-hmC) were found to be linked with epigenetic reprogramming.The present study aimed to examine the expression of the TET family and 5-hmC in the villi of human embryos and compared their expression between normal pregnancy and early pregnancy loss (EPL).Embryonic villi were collected from normal pregnant women (control) experiencing medical abortion and from EPL patients at gestation ages of 6,7 and 8 weeks.The mRNAs of TET family were analysed using quantitative polymerase chain reaction (qPCR),and TET proteins using Western blotting and immunohistochemical analysis.The MethylFlashTM Kit was used to quantify the absolute amount of 5-methylcytosine (5-mC) and 5-hmC.Our results showed that the expression of the TETs and 5-hmC in the normal villus decreased with increasing gestational age.Immunohistochemistry revealed that the TET proteins were expressed in the cytoplasm of trophoblasts and their expression was the highest in the 6-week tissue samples,which was consistent with the qPCR and Western blot results.The expression of TET1,TET2,and TET3 was lower in the villi in EPL group than in normal pregnancy group (P＜0.05 for all).It was concluded that the TET family and 5-hmC are critical in epigenetic reprogramming of human embryo.The findings also suggest that a deficiency of TETs in the villus might be associated with human EPL.

Effect of cottonseed oil and ferrous sulfate in broiler diets on intestinal morphometry at 42 days of age / Efeito de óleo de semente de algodão e sulfato ferroso em dietas de aves na morfometria intestinal aos 42 dias de idade

The aim of the study was to evaluate the intestinal morphometry of broilers at 42 days of age fed diets containing different levels of cottonseed oil, with and without addition of ferrous sulfate. A total of 560 male Ross chickens were used in a completely randomized design with a 4×2 factorial arrangement and five replications with 14 birds. The cottonseed oil levels tested were 0%, 2%, 4%, and 6%. At 42 days of age, one bird per replicate was collected for morphometric. Analyzed variables were villus height, crypt length and muscle wall thickness. The statistical analysis was performed using SAS, P=0.05. In the duodenum and ileum, there was no significant effect of interaction between the treatments. In the jejunum, only the length of the crypt, the treatment with cottonseed oil 2%, more ferrous sulfate resulted in lower values; however, the same oil content without ferrous sulfate led to a greater length, which was also observed in the treatment with 4% oil plus ferrous sulfate. Inclusion of cottonseed oil in broiler diets at 42 days of age does not impair their intestinal morphology. Supplementation with ferrous sulfate at these levels is unnecessary, since no improvement was observed in the intestinal morphometric.(AU)

O objetivo de estudo foi avaliar a morfometria intestinal de aves aos 42 dias de vida alimentadas com dietas contendo níveis variados de óleo de semente de algodão com e sem adição de sulfato ferroso. Um total de 560 machos de aves Ross foram usadas em estudo randomizado com amostras 4x2 e 5 replicações com 14 aves. Os níveis de óleo de semente de algodão testados foram 0%, 2%, 4%, e 6%. Aos 42 dias de idade, um pássaro por replica foi coletado para morfometria. As variáveis analisadas foram altura de villus, comprimento de cripta, e espessura de parede muscular. A análise estatística foi realizada com SAS, P=0,05. Não houve efeito significativo de interação entre tratamentos no duodeno ou íleo. No jejuno, apenas o comprimento da cripta, no tratamento com óleo de semente de algodão a 2% com mais sulfato ferroso, apresentou valor mais baixo. No entanto, o mesmo valor de óleo sem sulfato ferroso resultou em maior comprimento, resultado percebido também com 4% de óleo. Inclusão de óleo de semente de algodão em dietas de aves aos 42 dias de idade não causa danos à morfologia intestinal. Suplementação com sulfato ferroso não é necessário, já que não houve melhora em morfometria intestinal.

Application of Next-Generation Sequencing in Detection of Mutation Gene in ZMPSTE24 in Prenatal Diagnosisa Chinese Pedigree with Pathological Chorioamniotic Membrane Separation / 中山大学学报(医学科学版)

[Objective] To describe a case of a rare,novel mutation causing recurrent chorioamniotic membrane separation in a Chinese family with combined next-generation sequencing (NGS) and Sanger sequencing.[Methods] For the affected fetus,potential mutation were detected by the conbinedcombined next-generation sequencing (NGS) and Sanger sequencing.And the prenatal diagnosis were identified by Sanger sequencing.[Results] A frameshifting mutation c.1389_1390delAG (inherited from mother),and a missense mutationc.1006 G ＞ C (inherited from mother) have been identified in the affected fetus (the second pregnancy).The prenatal diagnosis of the third fetus turns out to be a carrier,the mutation was inherited from father.[Conclusions] We describe a novel mutation in gene ZMPSTE24,which was considered with mandibuloacral dysplasia with type B,and that may be the cousecoursecausing of recurrent chorioamniotic membrane separation.This rare mutation constitutes an additional heterogeneous defect causing chorioamniotic membrane separation.And the conbinedcombined next-generation sequencing (NGS) and Sanger sequencing allows high resolution characterization of novel mutions that are not readily detected by present methods.

Application of CNVplex combined with STR assay for genetic etiology exploration in chorionic villus of recurrent spontaneous abortion / 重庆医学

Objective The study is to present a novel assay for rapid detection of fetal aneuploidies in chorionic villus for spontaneous abortion.Methods Fetal chorionic villus samples were collected from 60 cases of women diagnosed with recurrent spontaneous abortion (RSA) before 13 weeks gestation.All samples were analyzed using CNVplex (copy numbcr variations multiplex) assay and fluorescence in situ hybridization (FISH) in addition to chromosome analysis.All villi specimens were cell cultured and karyotyped to confirm the fetal chromosomal status.Results Among 48 successfully cultured and karyotyped samples,the chromosomal abnormality rate was 60.42 ％.The results of karyotyping and the CNVplex assay were identical,both yielding 20 cases of euploidies,23 autosomal aneuploidies,3 triplodies and 2 × monosomies(Tumer Syndrome).However,FISH obtained only 38 results identical to karyotyping.Two cases of deletion and duplication of chromosome were also identified by CNVplex but not always by karyotyping.As for non-mosaic and non structural abnormity samples,the concordance between cytogenetics and genoty ping was 100％ in CNVplex and 79.17％ in FISH.Conclusion With CNVplex combined with STR(short tandem repeat) assay,we can detect the aneuploidy abnormalities as effectively as routine karyotyping without the need for cell culture,while also analyzing deletions and duplications(larger than 5 Mbp) that are not always detected by karyotype analysis.Our study demonstrates that CNVplex assay is an efficient,convenient,and accurate method to explore the etiology of miscarriage.

Changes of small intestinal villi microcirculation in sidestream dark-field imaging with different target blood pressure in rabbits during endotoxin shock / 中华危重病急救医学

Objective Changes of small intestine villus microcirculation perfusion in sidestream dark-field (SDF) imaging in the rabbits during endotoxic shock after fluid resuscitation with different target mean arterial pressure (MAP), and evaluation of feasibility of monitoring small intestine villus microcirculation by SDF were studied. Methods Sixty standard New Zealand white rabbits were randomly divided into two groups: low target MAP group (group A, n = 30) and high target MAP group (group B,n = 30). Fistula operation of ileum was madein vitro, and lipopolysaccharide (LPS, 2 mg/kg) was injected to establish endotoxic shock model. Group A was administered with the lower dose fluid resuscitation (lactated Ringer solution, 20 mL·kg-1·h-1) for target MAP of 65 mmHg (1 mmHg =0.133 kPa); group B was administered with the higher dose fluid resuscitation (lactated Ringer solution, 30 mL·kg-1·h-1) for MAP of 80 mmHg. Continuous norepinephrine intravenous injection (0.5-1.0μg·kg-1·min-1) was administered only after fluid therapy couldn't reach the target MAP. The changes of small intestine villus microcirculation perfusion indexes such as vessels per villus (VV), proportion of perfused villi (PPV), microvascular flow index (MFI), borders of villus score (BVS), vessels villus score (VVS) were continuously observed and recorded before the shock, during the shock and after fluid resuscitation using SDF imaging. The differences of microcirculation perfusion were compared between two groups using the specific parameter evaluation system to determine severity of villi microcirculation and injury scores at different stages.Results VV and borders of villus were clear and contact before shock in two groups. After shock, VV, PPV were significantly decreased in both two groups, the borders of villus were destroyed, MFI, BVS, VVS and the total score of villi injury microcirculation were obviously and severely decreased. Partial blood flow of villous capillaries after fluid resuscitation was recovered in two groups, but the perfusion of some region was un-balanced with the outworn borders of villus. VV were rose as compared before and after fluid resuscitation in groups A and B (vessels: 1.21±0.22 vs. 0.81±0.12, 1.54±0.28 vs. 0.79±0.13), and PPV [(31±4)% vs. (12±2)%, (38±5)% vs. (13±3)%], MFI (1.55±0.09 vs. 1.09±0.03, 1.97±0.11 vs. 1.05±0.03), VVS (points: 1.22±0.08 vs. 0.89±0.02, 2.06±0.15 vs. 0.90±0.02) and the sum of MFI, BVS, VVS (3.70±0.19 vs. 2.85±0.07, 5.01±0.29 vs. 2.88±0.08) were significant rose (allP 0.05).Conclusions For the small intestine villus microcirculation perfusion, the higher target MAP (80 mmHg) after fluid resuscitation or/and vasoconstrictor drugs usage were probably better than the relatively lower target MAP (65 mmHg) during endotoxic shock. SDF imaging is a very promising technique for intestinal villi microcirculatory visualization and assessment.

Clinical application of high-throughput gene sequencing technology and STR in chromosome karyotype analysis of the villus tissues of spontaneous abortion of spontaneous abortion / 中国综合临床

Objective To analyze the clinical application of high?throughput gene sequencing technolo?gy and STR in chromosome karyotype analysis of the villus tissues of spontaneous abortion. Methods In 27 ca?ses of spontaneous abortion after pregnancy,classic cell of villus tissues culture and chromosomal karyotype anal?ysis,and high ?throughput gene sequencing technology and STR were performed,and then compared the analysis results of two methods. Results ( 1) The successful rate of cell of villus tissues culture and chromosomal karyo?type analysis was 85%( 23/27) ,of high?throughput gene sequencing technology and STR was 96%( 26/27) ,and the difference was not significant( P>0. 05) ( 2) In the 4 cases that failed in karyotype analysis,there were 3 ca?ses showed abnormal chromosomal number variation( CNV) in high?throughput gene sequencing technology and STR. ( 3) Of the 23 cases,chorionic villus was successfully cultured in 10 cases,abnormal karyotypes were iden?tified in 13 cases,the positive rate was 57%. Of the 26 cases,high?throughput gene sequencing technology and STR was successfully checked in 3 cases,abnormal CNV were identified in 23 cases,the positive rate was 85%, the difference was significant(χ2=6.387,P<0.05). (4)The rates of chromosomal number abnormality were 52%( 12/23) and 50% ( 13/26) of karyotype analysis and chromosome aberration detection,respectively. In 10 cases of normal cell culture karyotype,there were 7 cases in the presence of micro deletion / micro repetition de?tected by high?throughput gene sequencing technology and STR. Conclusion The method of massively parallel sequencing in chromosome analysis,compared with the method of cell of villus tissues culture and chromosome a?nalysis,can be accurate and quick,and has high successful rate in detecting the chromosome of non aneuploid and deletion/duplication abnormality,which can be a good complementary and alternative method of the classic cell of villus tissues culture and chromosome karyotype analysis.

Dietas suplementadas com óleos essenciais não promove alterações na morfometria de vilos e criptas do duodeno de frangos de corte / Diets supplemented with essential oils do not promote alterations in villous morphometry and duodenum crypt of broilers / Dietas suplementadas con aceites esenciales no promove cambios en morfometría de las vellosidades y criptas del duodeno de pollos de engorda

Devido à necessidade da substituição dos antimicrobianos como melhoradores de desempenho por aditivos alternativos, inúmeros produtos estão em estudo. Dentre esses estão os óleos essenciais que apresentam características antimicrobianas, contribuindo para manutenção da integridade intestinal e consequentemente melhora no desempenho, garantindo a segurança alimentar, evitando a resistência cruzada, bem como a contaminação do meio ambiente. Dessa forma, objetivou-se avaliar os efeitos da inclusão de uma mistura de óleos essenciais extraídos de orégano, alho, limão, alecrim, timo, eucalipto e laranja doce, na dosagem de 300 mL para 500 L de água de bebida, sobre a morfometria da mucosa do duodeno de frangos de corte. Foram avaliados quatro aviários tratados com óleos essenciais na ração e quatro aviários controle, durante o período de um a sete dias de idade, contendo pintainhos da linhagem Cobb. Para a análise morfométrica, cinco aves de cada aviário (total de 40 aves) foram mortas, utilizando-se de protocolo anestésico, medicação pré-anestésica de xilazina 4mg/Kg IM. e após solução de tiopental na dose de 25 mg/Kg IV. Após eutanásia, foram colhidas amostras de material do intestino delgado, correspondente ao segmento duodenal, para realização das medições de altura de vilo e profundidade de cripta. Não foram verificados efeitos sobre a altura de vilo e profundidade de cripta nos animais tratados com óleos essenciais (P>0,05). Nas condições em que foram realizados o experimento, pode-se concluir que os óleos essenciais não alteraram a altura de vilo e a profundidade de cripta do duodeno.

Due to the need for replacement of antimicrobials as performance enhancers to alternative additives, several products are being studied. Among them, essential oils that exhibit antimicrobial properties are being analyzed, which contribute to the maintenance of intestinal integrity and consequently improving performance, assuring food safety, preventing cross-resistance, as well as contamination of the environment. Thus, the objective of this study is to evaluate the effects of adding a mixture of essential oils from oregano, garlic, lemon, rosemary, thyme, eucalyptus and sweet orange, at a dosage of 300 mL for 500 L of drinking water, on the morphometry of duodenal mucosa of broilers. Four poultry aviaries treated with essential oils in the diet and four control poultry aviaries were evaluated during the period from one to seven day old, containing Cobb chicks. For the morphometric analysis, five birds in each aviary (total of 40 birds) were euthanized using anesthetic protocol premedication with xylazine 4mg/kg IM) and after, a solution of thiopental (25 mg /Kg IV). After euthanasia, material samples were collected from the small intestine (duodenum) for measuring villus height and crypt depth. No effects on villus height and crypt depth could be found in the animals treated with essential oils (P> 0:05). Under the experimental conditions, it can be concluded that essential oils did not alter villus height and crypt depth of the duodenum.

Debido a la necesidad de reemplazo de antimicrobianos como mejoradores de desempeño por aditivos alternativos, innúmeros productos están siendo estudiados. Entre esos están los aceites esenciales que presentan características antimicrobianas, lo que contribuye al mantenimiento de la integridad intestinal y por consiguiente mejora el rendimiento, por garantizar la seguridad alimentaria, evitando resistencia cruzada, así como la contaminación del medio ambiente. Por lo tanto, el objetivo ha sido evaluar los efectos de la adición de una mezcla de aceites esenciales extraídos del orégano, ajo, limón, romero, tomillo, eucalipto y naranja dulce, a una dosis de 300 ml para 500 litros de agua potable, sobre la morfometría de la mucosa duodenal de pollos de engorde. Se han evaluado cuatro aviarios tratados con aceites esenciales en el pienso y cuatro aviarios control, durante el período de uno a siete días de edad, conteniendo pollitos Cobb. Para el análisis morfométrico, cinco aves de cada aviario (total de 40 aves) fueron sacrificadas, utilizándose de protocolo anestésico, medicación pre anestésica de xilazina 4 mg /Kg IM., y después tiopental solución a una dosis de 25 mg /Kg IV. Después de la eutanasia, se ha recogido muestras de material del intestino delgado, que corresponde al segmento duodenal, para llevar a cabo las mediciones de la altura de las vellosidades y profundidad de la cripta. No se ha verificado efectos sobre la altura de las vellosidades y la profundidad de la cripta en los animales tratados con aceites esenciales (P> 0,05). En las condiciones en que se han realizado el experimento, se puede concluir que los aceites esenciales no alteraron la altura de las vellosidades y la profundidad de la cripta del duodeno.

Effects of Solanine on the human colon cancer in chicken embryo transplantation model angiogenesis / 重庆医学

Objective To establish chicken embryo transplantation model of human colon cancer and to research the effect of so‐lanine on angiogenesis .Methods Cases with chicken embryos were divided into the low‐,mid‐and high dose solanine group and con‐trol group ,with 10 cases in each groups ,and then the cultured human colon cancer cell line HT‐29 cell lines were inoculated to the chicken embryo villus allantois membrane (CAM ) .We observed the characteristics of the transplanted tumor in CAM angiogenesis by the stereo microscope .Image analysis software of Image‐pro plus 6 .0 and immunohistochemical method were used to observe the effect of different dose of solanine on angiogenesis .Results HT‐29 cell lines were inoculated to CAM 3-5 days ,a large number of blood vessels concentrated in tumors ,growing into or acrossing the surface of tumors .While tumors also rapidly growed .We took photo on the 5th day after receiving medicine and did imaging analysis .Then we calculated the area of angiogenesis in experimental group ,which was significantly lower than that of the control group ,quantitatively in a dose‐dependent manner .There were signifi‐cant differences among the groups(P<0 .01) .Microvascular density of 3 different dose of solanine was significantly lower than that of the control group by immunohistochemical method ;the expression of Ki‐67 antigen index decreased gradually ,which was highest in the control group ,and there were significant differences among the groups (P<0 .01) .Conclusion Solanine could inhibit angio‐genesis induced by human colon cancer HT‐29 cell lines obviously ,thus inhibiting the growth of tumor and providing an important basis for the treatment of anti‐tumor angiogenesis .

Analysis of fetal chromosomal karyotypes in 1 160 pregnant women during the first trimester of gestation / 重庆医学

Objective To evaluate the value of chorionic villus cells karyotype analysis in prenatal diagnosis during the first tri-mester of pregnancy.Methods Pregnant women with prenatal diagnosis indications were punctured by guiding abdominal B-mode ultrasound to get villi tissue which was then to develop cell culture,chromosome preparation and karyotype analysis.Results A to-tal of 1 140 cases were successfully cultured,and the successful cultivating rate was 98.2% (1 140/1 160).Among them,chromo-somes of 62 cases were detected to be non-polymorphic structural abnormalities,including 32 abnormal chromosome number,5 chro-mosome balanced translocation,3 chromosome deletion,and 22 chimeras.What′s more,20 cases were detected to be chromosomal inversion,19 cases of chromosome 9 were inversion,and one with chromosome Y was inversion.Conclusion Karyotype analysis of villus cell could help to detect fetal chromosomal abnormalities during early pregnancy and get early intervention.It was significant to reduce the child′s birth with chromosome abnormalities.

Chorionic villus sampling

Chorionic villus sampling has gained importance as a tool for early cytogenetic diagnosis with a shift toward first trimester screening. First trimester screening using nuchal translucency and biomarkers is effective for screening. Chorionic villus sampling generally is performed at 10-12 weeks by either the transcervical or transabdominal approach. There are two methods of analysis; the direct method and the culture method. While the direct method may prevent maternal cell contamination, the culture method may be more representative of the true fetal karyotype. There is a concern for mosaicism which occurs in approximately 1% of cases, and mosaic results require genetic counseling and follow-up amniocentesis or fetal blood sampling. In terms of complications, procedure-related pregnancy loss rates may be the same as those for amniocentesis when undertaken in experienced centers. When the procedure is performed after 9 weeks gestation, the risk of limb reduction is not greater than the risk in the general population. At present, chorionic villus sampling is the gold standard method for early fetal karyotyping; however, we anticipate that improvements in noninvasive prenatal testing methods, such as cell free fetal DNA testing, will reduce the need for invasive procedures in the near future.

The effect of interferon alfα-2b on the ultrastructure and Caspase-3 levels in villus in early pregnancy with bacterial vaginal disease / 实用医学杂志

Objective To investigate the effect of interferon alfα-2b on the ultrastructure and Caspase-3 levels in villus in early pregnancy with bacterial vaginal disease (BV). Method Early pregnant women were divided into two groups. The treated group included 25 early pregnant women with BV who chose to have an early termination and were treated with rhINFα-2b. The controling group included 30 early pregnant women without any genital tract infectious diseases. The caspase-3 levels in trophocytes were detected by immunochemistry and the ultrastructural changes were observed in villus by transmission electron microscopy. Result (1)There was no apparent difference of ultrastructural changes between the two groups. (2)There was no statistical significance of the levels of caspase-3 between the two groups (P>0.05). Conclusion The excessive apoptosis do not occur in the trophocytes when treated with INFα-2b.

Villus cell culture and its karyotype analysis for 112 women who had spontaneous abortion in early pregnancy / 国际检验医学杂志

Objective To investigate the clinical relationship between villus cells chromosome abnormality and spontaneous a-bortion in early pregnancy.Methods Villus cells from 112 women who just had spontaneous abortion in early pregnancy were cul-tured and prepared for karyotype analysis.Results 105 specimens were anaIyzed successfully which accounted for 93.75%(105/112).Among them 50 specimens were normal karyotype which accounted for 47.62%(50/105),55 cases were abnormal karyotype accounted for 52.38%(55/105).Abnormal number was the major type of abnormal karyotype.Among them there were 33 cases of trisomy which were most common,6 cases of triploid,1 case of tetraploid and 1 case of compound,2 cases of chimera,1 case of sex-ual chromatids and 6 cases of structural abnormalities.In the specimens with normal karyotype,20 cases were from male,30 cases were from female.Conclusion Embryo chromosome abnormality is one of the factors causing early pregnancy spontaneous abor-tion.Conduct villus chromosome examination in clinical treatment is useful to determine the reasons of abortion and to guide the next pregnancy.

Efeitos do espaço de alojamento e da idade sobre desenvolvimento, integridade e proteção da mucosa intestinal de frangos de corte / Effects of enclosure size and age on development, integrity and protection of the intestinal mucosa in broiler chickens

Foi avaliado se o espaço de alojamento influencia o desenvolvimento da mucosa do intestino delgado de frangos, sendo usado um esquema fatorial 4 x 2 (14, 28, 42, 55 dias de idade e alojamento pequeno: 0,11m²/ave e grande: 0,525m²/ave). Foram analisadas: altura, perímetro e número dos vilos, profundidade das criptas, número de células caliciformes e número de vilos sem e com perda de epitélio e exposição do conjuntivo. Em ambos os espaços de alojamento, o número de vilos diminuiu e a profundidade das criptas aumentou com a idade nas três regiões intestinais. A altura e o perímetro dos vilos aumentaram de 14 para 28 dias no jejuno e íleo, e o número de células caliciformes foi maior no duodeno e íleo a partir dos 42 dias. A perda de epitélio pelos vilos do íleo aumentou com a idade. Nas aves do alojamento grande, ocorreu aumento na altura e no perímetro dos vilos duodenais com a idade, possibilitado pelo aumento na profundidade das criptas, que resultou em vilos maiores nessas aves do que nas do alojamento pequeno aos 55 dias de idade. O número de células caliciformes diminuiu nas aves do alojamento pequeno aos 42 dias de idade. Os dados mostram que o tamanho do alojamento interferiu no modelo de desenvolvimento temporal da mucosa intestinal e que frangos apresentaram maior tamanho de vilo duodenal quando alojados em espaço maior.

This study evaluated if enclosure size and age influence the intestinal mucosa development of broiler chickens, using an experimental design in an 4x2 factorial arrangement (14, 18, 42 and 55 days of age and two enclosure spaces: small= 0.11m² /bird and large= 0.525m²/bird). Villous height, perimeter and number, and crypt depth, goblet cells number, and number of villous without and with epithelium loss and conjunctive tissue exposition were analyzed in duodenum, jejunum and ileum. Regardless of enclosure size, the villous number diminished and the crypt depth increased with the age in the three intestinal regions. From 14 to 28 days of age, villous height and perimeter increased in the jejunum and ileum. The number of goblet cells was greater from 42 days in duodenum and ileum, and epithelium loss increased until 42 days in the ileum. Broilers housed in small and large enclosures differed in the intestinal characteristics from 42 days. In broilers housed in large enclosure, duodenal villous height and perimeter increased with the age, with an increase in the crypt depth, reaching greater size at 55 days of age than duodenal villous of the broilers housed in a small enclosure. These later presented at 42 days a smaller number of goblet cells than the former. According these results, enclosure size influenced the temporal pattern of the intestinal mucosa development, and broilers presented major duodenal villous growth when housed in large enclosures.

Paracentric Inversions Found in Prenatal Diagnosis

PURPOSE: This study was designed to confirm whether the paracentric inversions of fetuses and parents may be harmless. MATERIALS AND METHODS: We report 10 cases (0.14%) with paracentric inversions among 7,181 prenatal cases observed during prenatal diagnosis performed at Cheil General Hospital between January 2009 and June 2013. We used cytogenetic GTL- and RBG-banding techniques. RESULTS: Of the 10 cases, nine cases were transmitted from each of the parents, and one case was de novo. Nine cases were phenotypically normal up to one month of age after birth. One case was lost to follow-up. We present prenatal diagnosis and follow-up examination of the fetuses with paracentric inversion. CONCLUSION: Based on our cases, most paracentric inversions are considered to be harmless. The precise identification of paracentric inversions might be clinically important and helpful for genetic counseling.

Análise dos resultados maternos e fetais dos procedimentos invasivos genéticos fetais: um estudo exploratório em Hospital Universitário / Analysis of fetal and maternal results from fetal genetic invasive procedures: an exploratory study at a University Hospital

OBJETIVO: Caracterizar as indicações das gestantes que procuraram o serviço de Medicina Fetal do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo para realização de procedimentos invasivos diagnósticos e avaliar os resultados dos cariótipos fetais e de suas gestações. MÉTODOS: Estudo observacional retrospectivo das gestantes que realizaram biópsia de vilo corial (BVC), amniocentese e cordocentese no período de fevereiro de 2005 a dezembro de 2009. Não foram incluídos outros procedimentos diagnósticos ou procedimentos terapêuticos. O resultado da gestação foi obtido através de consulta de prontuário eletrônico e/ou físico e/ou contato telefônico. RESULTADOS: Foram realizados 713 procedimentos (113 BVC, 340 amniocenteses e 260 cordocenteses). A principal indicação para a realização dos procedimentos invasivos foi a presença de alterações estruturais nos fetos, seguido por valores aumentados da translucência nucal e pela idade materna avançada. O cariótipo fetal esteve alterado em 186 casos (26,1%). A trissomia do cromossomo 18 foi a aneuploidia mais comum, seguida pela trissomia do 21, a monossomia do X e a trissomia do cromossomo 13. Ocorreram 4,9% de abortamento, 25,7% de natimortos e 13% de neomortos. Oito gestantes optaram pela interrupção judicial, e 99% das gestantes cujos fetos não apresentavam malformação e que apresentavam cariótipo fetal normal tiveram nativivos.

OBJECTIVE: To characterize the indications of pregnant women who sought the Fetal Medicine Services of the Hospital das Clínicas, at the Medical School of the Universidade de São Paulo for performing invasive diagnostic procedures, and to evaluate the results of fetal karyotypes and their pregnancies. METHODS: A retrospective and observational study on pregnant women who underwent chorionic villus sampling (CVS), amniocentesis, and cordocentesis in the period from February, 2005 to December, 2009. Other diagnostic or therapeutic procedures were not included. The result of pregnancy was obtained by consulting patient electronic records, medical records, and/or telephone call. RESULTS: 713 procedures were performed (113 CVS, 340 amniocenteses, and 260 cordocenteses). The main indication for performing invasive procedures was the presence of structural changes in fetuses, followed by increased values of nuchal translucency, and advanced maternal age. Fetal karyotype was altered in 186 cases (26.1%). The 18 trisomy was the commonest aneuploidy followed by the 21 trisomy, X monosomy, and 13 trisomy. There were 4.9% cases of miscarriage, 25.7% cases of stillborn infants, and 13% cases of neonatal deaths. Eight pregnant women opted for legally induced abortion. 99% of pregnant women whose fetuses did not present abnormalities and presented normal fetal karyotype had infants who were born alive.

Effects of starter diet supplementation with arginine on broiler production performance and on small intestine morphometry / Efeito da suplementação de arginina na dieta de frangos na performance produtiva e na morfologia do intestino delgado

The effects of starter diet (days 1 to 21) supplemented with arginine (Arg) on the production performance and duodenum and jejunum mucosa morphometry of broilers were studied. Male Cobb broiler chickens (990) were randomly assigned to one of five treatments in a complete random design. Measurements of 33 chicks per treatment were made in six repetitions. The treatments consisted of a basal diet with 1.390% digestible Arg (no supplementation) and four dietary levels (1.490%, 1.590%, 1.690%, and 1.790%), providing a relationship with lysine of 1.103; 1.183; 1.262; 1.341 and 1.421%, respectively. From the age of 22 days on, all birds received conventional grower diet. The data were submitted to regression analysis by polynomial decomposition of the degrees of freedom in relation to the levels of Arg. The Arg supplementation increased (P<0.05) the live weight and the feed conversion ratio without increasing the feed intake of the birds. However, no effect was observed (P>0.05) in the growth phase (days 22 to 42) in the absence of the Arg supplementation. The supplementation of Arg over of NRC recommendation during the starter phase may be necessary for the expression of the maximal weight gain potential in birds. No effect (P<0.05) of Arg dietary supplementation was observed either on small intestine weight and length at any age. However, the duodenum villus:crypt ratio increased and the crypt depth decreased in the first week in response to increasing dietary Arg. It is concluded that broiler Arg dietary supplementation in the starter diet improved production performance and small intestine morphometry, especially in the first week.

O efeito na dieta inicial (1-21 dias) da suplementação de arginina (Arg) foi estudada sobre o desempenho e a morfologia da mucosa do jejuno em frangos de corte. Pintos machos Cobb (990) foram utilizados em um delineamento inteiramente casualizado com 5 tratamentos e 6 repetições com 33 aves cada. Os tratamentos consistiram em uma dieta basal com 1,390% de Arg digestível (sem suplementação) e 4 dietas com adição (1,490%; 1,590%; 1,690% e 1,790%), fornecendo uma relação de lisina de 1,103; 1,183; 1,262; 1,341 e 1,421%, respectivamente. A partir de 22 dias todas as aves receberam ração convencional. Os dados foram submetidos à análise de regressão pela decomposição polinomial dos graus de liberdade, referentes aos níveis de Arg A suplementação de Arg melhorou (P<0,05) o peso vivo e a conversão alimentar sem aumentar o consumo de ração. No entanto, não houve efeito (P>0,05) na fase de crescimento (22 a 42 dias) na ausência de suplementação de Arg. A suplementação acima do recomendado pelo NRC na fase inicial pode ser necessária para a expressão máxima do potencial de ganho de peso em aves. Não houve efeito da suplementação de Arg na dieta no peso e comprimento do intestino delgado em nenhuma idade. No entanto, a relação vilo:cripta no duodeno aumentou e a profundidade da cripta diminuiu na primeira semana em resposta ao incremento de Arg na dieta. Concluiu-se que em frangos de corte a suplementação de Arg na dieta inicial melhorou o desempenho e a morfometria do intestino delgado, especialmente na primeira semana.