Impact of inactivated SARS-CoV-2 vaccination on embryo ploidy: a retrospective cohort study of 133 PGT-A cycles in China

BACKGROUND: Unsubstantiated concerns have been raised on the potential correlation between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination and infertility, leading to vaccine hesitancy in reproductive-aged population. Herein, we aim to evaluate the impact of inactivated SARS-CoV-2 vaccination on embryo ploidy, which is a critical indicator for embryo quality and pregnancy chance. METHODS: This was a retrospective cohort study of 133 patients who underwent preimplantation genetic testing for aneuploidy (PGT-A) cycles with next-generation sequencing technology from June 1st 2021 to March 17th 2022 at a tertiary-care medical center in China. Women fully vaccinated with two doses of Sinopharm or Sinovac inactivated vaccines (n = 66) were compared with unvaccinated women (n = 67). The primary outcome was the euploidy rate per cycle. Multivariate linear and logistic regression analyses were performed to adjust for potential confounders. RESULTS: The euploidy rate was similar between vaccinated and unvaccinated groups (23.2 ± 24.6% vs. 22.6 ± 25.9%, P = 0.768), with an adjusted ß of 0.01 (95% confidence interval [CI]: -0.08-0.10). After frozen-thawed single euploid blastocyst transfer, the two groups were also comparable in clinical pregnancy rate (75.0% vs. 60.0%, P = 0.289), with an adjusted odds ratio of 6.21 (95% CI: 0.76-50.88). No significant associations were observed between vaccination and cycle characteristics or other laboratory and pregnancy outcomes. CONCLUSIONS: Inactivated SARS-CoV-2 vaccination had no detrimental impact on embryo ploidy during in vitro fertilization treatment. Our finding provides further reassurance for vaccinated women who are planning to conceive. Future prospective cohort studies with larger datasets and longer follow-up are needed to confirm the conclusion.

Yield, structural composition and nutritive characteristics of ryegrass cultivars used to haymaking in lowland soils / Rendimento, composição estrutural e características nutritivas de cultivares de azevém utilizadas para fenação em solos de várzea

This research aimed to evaluate the dry matter yield, structural composition and nutritive characteristics of diploid and tetraploid annual ryegrass cultivars on different phenological development to haymaking on lowland soils. The experimental design was developed based on randomized blocks with split plots, four cultivars of annual ryegrass (BRS Ponteio and FEPAGRO São Gabriel, diploid; INIA Escorpio and KLM 138, tetraploid), and three phenological crop phases (vegetative, pre-blossoming and blossoming). Were evaluated, dry matter yield, leaf:stem ratio, leaf weight ratio, tiller population density, specific leaf area, crude protein, neutral detergent fiber, and acid detergent fiber. All data were submitted to analysis of variance and the means were compared by Tukey-Kramer test (p< 0,05). Highest forage mass is obtained with harvest in blossoming stage. Tetraploid cultivars present better leaf proportion and higher content of crude protein during vegetative stage. The decrease the in concentration of protein with the change of phenological stage is less evident on diploid cultivars. The vegetative stage enables harvest forage with high nutritional value, with lower production of biomass.

O objetivo do trabalho foi avaliar o rendimento de matéria seca, composição estrutural e características nutritivas de cultivares de azevém anual, diploides e tetraploides em diferentes estádios fenológicos, para fenação, em solos de várzea. O delineamento experimental foi de blocos ao acaso, com parcelas divididas, com quatro cultivares de azevém anual (BRS Ponteio e FEPAGRO São Gabriel, diploides; INIA Escorpio e KLM 138, tetraploides) e três estádios fenológicos (vegetativo, pré-florescimento e florescimento). Foram avaliadas: rendimento de matéria seca, relação folha/colmo, razão de peso foliar, densidade populacional de perfilhos, área foliar específica, proteína bruta, fibra em detergente neutro e fibra em detergente ácido. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste Tukey-Kramer (p <0,05). A maior massa de forragem é obtida com a colheita no estádio de florescimento. As cultivares tetraploides apresentam melhor proporção foliar e maior teor de proteína bruta durante o período vegetativo. A diminuição na concentração de proteína, com a mudança de estádio fenológico é menos evidente nas cultivares diploides. O estágio vegetativo possibilita a colheita de forragem com alto valor nutricional, mas com menor produção de biomassa.

Karyotype analysis in Bignonieae (Bignoniaceae): chromosome numbers and heterochromatin

ABSTRACT Chromosome numbers and heterochromatin banding pattern variability have been shown to be useful for taxonomic and evolutionary studies of different plant taxa. Bignonieae is the largest tribe of Bignoniaceae, composed mostly by woody climber species whose taxonomies are quite complicated. We reviewed and added new data concerning chromosome numbers in Bignonieae and performed the first analyses of heterochromatin banding patterns in that tribe based on the fluorochromes chromomycin A3 (CMA) and 4'-6-diamidino-2-phenylindole (DAPI). We confirmed the predominant diploid number 2n = 40, as well as variations reported in the literature (dysploidy in Mansoa [2n = 38] and polyploidy in Dolichandra ungis-cati [2n = 80] and Pyrostegia venusta [2n = 80]). We also found a new cytotype for the genus Anemopaegma (Anemopaegma citrinum, 2n = 60) and provide the first chromosome counts for five species (Adenocalymma divaricatum, Amphilophium scabriusculum, Fridericia limae, F. subverticillata, and Xylophragma myrianthum). Heterochromatin analyses revealed only GC-rich regions, with six different arrangements of those bands. The A-type (one large and distal telomeric band) were the most common, although the presence and combinations of the other types appear to be the most promising for taxonomic studies.

Pathogenic variant in NLRP7 (19q13.42) associated with recurrent gestational trophoblastic disease: Data from early embryo development observed during in vitro fertilization / 대한생식의학회지

OBJECTIVE: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. METHODS: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. RESULTS: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. CONCLUSION: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.

Utility of p57KIP2 and Her-2 fluorescence in situ hybridization in differentiating partial from complete hydatidiform mole

INTRODUCTION: Hydatidiform mole (HM) is an abnormal gestation characterized by significant hydropic enlargement, trophoblastic proliferation and atypia involving part or all of the chorionic villi. The diagnosis and classification of hydatidiform moles is subject to great inter-observer variability due to significant morphologic overlaps. This study aims to evaluate the utility of p57KIP2 immunohistochemistry and ploidy by Her-2 FISH in refining the diagnosis of molar tissues. METHOD: 113 and 78 molar cases were retrieved from the archives of the Histopathology Section of the Philippine General Hospital and Pathology Department of Seoul National University Hospital, respectively. TMA sections were submitted for immunohistochemical analysis for p57KIP2. Ploidy was determined by fluorescence in situ hybridization using Her-2 probe. An interrater reliability analysis was done using the Kappa statistics with 95% confidence interval. RESULTS: All 68 (100%) cases diagnosed as CH were negative for p57KIP2 staining and are diploid. Among the 54 cases of PH, only 1 (2%) is positive for p57KIP2 and is diploid. The interrater reliability between p57KIP2 and Her-2 FISH ploidy results is 0.66 (p CONCLUSION: Morphologic assessment alone may not be sufficient in problematic cases. p57KIP2 in conjunction with by Her-2 FISH are good adjuncts in the diagnosis and classification of hydatidiform mole.

Clinical Value of Immunophenotype Detection and DNA Ploid Analysis by Flow Cytometry for Non-Hodgkin's Lymphoma Patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical values of detecting immunophenotype and analyzing DNA ploid by flow cytometry for patients with non-Hogkin's lymphoma (NHL).</p><p><b>METHODS</b>Eighty NHL patients admitted in our hospital from August 2007 years to March 2015 Years were included in the observation group, 20 patients with reactive lymphoid hyperplasias were selectod as control group. The immunophenotypes were detected and the DNA ploid was analyzed by flow cytometry.</p><p><b>RESULTS</b>The detected rate of DNA aneuploidy, DAN index (DI) and SPF in observation group were higher than those in control group, there was signifificant difference (P < 0.05). The SPF and DI in patients with NHL-I, NHL-II had no statistical difference as compared with that in control group (P > 0.05); but the SPF and DI in pateints with NHL-III and patients with NHL-IV showed statistical significance as compared with that in control group (P < 0.05). The SPF and DI in patients with low malignancy group and middle malignancy group showed statistical significance as compared with control group (P < 0.05). The SPF and DI in middle malignancy group had statistical significance as compared with that in low malignancy group (P < 0.05).</p><p><b>CONCLUSION</b>the immunophenotype detection and DNA ploid analysis by flow cytometry can reflect the tumor proliferation and deterioration of patients with Non-Hogkin's lymphoma, predicting the prognosis.</p>

A retrospective analysis of cytogenetic alterations in patients with newly diagnosed multiple myeloma: a single center study in Korea

BACKGROUND: The accurate identification of cytogenetic abnormalities in multiple myeloma (MM) has become more important over recent years for the development of new diagnostic and prognostic markers. In this study, we retrospectively analyzed the cytogenetic aberrations in MM cases as an initial assessment in a single institute. METHODS: We reviewed the cytogenetic results from 222 patients who were newly diagnosed with MM between January 2000 and December 2015. Chromosomal analysis was performed on cultured bone marrow samples by standard G-banding technique. At least 20 metaphase cells were analyzed for karyotyping. RESULTS: Clonal chromosome abnormalities were detected in 45.0% (100/222) of the patients. Among these results, 80 cases (80.0%) had both numerical and structural chromosome abnormalities. Overall hyperdiploidy with structural cytogenetic aberrations was the most common finding (44.0%), followed by hypodiploidy with structural aberrations (28.0%). Amplification of the long arm of chromosome 1 and -13/del(13q) were the most frequent recurrent abnormalities, and were detected in 50 patients (50.0%) and 40 patients (40.0%) with clonal abnormalities, respectively. The most common abnormality involving 14q32 was t(11;14)(q13;q32), which was observed in 19 cases. CONCLUSION: These findings demonstrate that myeloma cells exhibit complex aberrations regardless of ploidy, even from a single center in Korea. Conventional cytogenetic analysis should be included in the initial diagnostic work-up for patients suspected of having MM.

The parental origin correlates with the karyotype of human embryos developing from tripronuclear zygotes / 대한생식의학회지

OBJECTIVE: It has previously been suggested that embryos developing from intracytoplasmic sperm-injected (ICSI) zygotes with three pronuclei (3PN) are endowed with a mechanism for self-correction of triploidy to diploidy. 3PN are also observed in zygotes after conventional in vitro fertilization (IVF). The parental origin, however, differs between the two fertilization methods. Whereas the vast majority of 3PN IVF zygotes are of dispermic origin and thus more likely to have two centrioles, the 3PN ICSI zygotes are digynic in origin and therefore, more likely to have one centriole. In the present study, we examine whether the parental origin of 3PN embryos correlates with the karyotype. METHODS: The karyotype of each nucleus was estimated using four sequential fluorescence in situ hybridizations-each with two probes-resulting in quantitative information of 8 different chromosomes. The karyotypes were then compared and correlated to the parental origin. RESULTS: 3PN ICSI embryos displayed a significantly larger and more coordinated reduction from the assumed initial 3 sets of chromosomes than 3PN IVF embryos. CONCLUSION: The differences in the parental origin-and hence the number of centrioles-between the 3PN IVF and the 3PN ICSI zygotes are likely to be the cause of the differences in karyotypes.

Lesão renal aguda como complicação da ingestão excessiva de suco do fruto biri biri (Averrhoa bilimbi)

O fruto biri-biri pertence à família das Oxalidacae, espécie Averrhoa bilimbi. Este fruto tem um alto conteúdo de oxalato solúvel e é utilizado na culinária, na produção de picles, geleias, e como tratamento para algumas doenças como hipertensão, diabetes e hiperlipidemia. Assim como outros frutos ricos em oxalato, pode provocar lesão renal aguda. Relatamos o caso de um paciente de 50 anos, hipertenso, com função renal normal, que ingeriu uma grande quantidade de suco em jejum para tratamento de hipertensão. O paciente desenvolveu quadro de lesão renal aguda associado a dores lombares, soluços e diarreia. A lesão renal aguda era não oligúrica e teve uma evolução favorável em 10 dias sem necessidade de tratamento dialítico. A função renal retornou ao normal após esses 10 dias de seguimento.

Sheep gastrointestinal helminthiasis in the Sertão region of Paraíba State, Northeastern Brazil: prevalence and risk factors / Helmintoses gastrintestinais de ovinos no Sertão do Estado da Paraíba, Nordeste do Brasil: prevalência e fatores de riscos

In this study, we aimed to establish the prevalence and risk factors relating to gastrointestinal helminthiasis, and to characterize the sanitary management practiced among sheep herds in the Sertão region of the state of Paraíba, northeastern Brazil, based on factors that condition the ways of controlling these parasites in these herds. The research was carried out between April and July 2012. We visited 54 farms, where fecal and blood samples were individually collected from 465 animals. On each farm, a questionnaire was applied to gather information on variables relating to potential risk factors. The prevalence of sheep gastrointestinal helminthiasis in the region was 75.9%. At least one animal tested positive for this helminthiasis on 53 (98.1%) of the 54 farms evaluated. The eggs per gram of feces (EPG) analysis showed the following infection burdens: 51.8% with mild infection, 27.1% moderate infection, 9.9% heavy infection and 11.2% fatal infection. Among the sheep farms visited, anthelmintics were used on 81.5% (p <0.05). The most relevant risk factor in this study was the farm area, because it defines the area available for grazing animals. Properties with many animals and little pasture area, which are the most abundant type in the Sertão region of Paraíba, tend to have high prevalence of gastrointestinal helminthiasis, because the animals are more prone to reinfection. The Sertão region of Paraíba presents high prevalence of gastrointestinal helminthiasis among sheep, and the farm area is the most relevant risk factor for the development of these parasites.

Objetivou-se determinar a prevalência e os fatores de risco para as helmintoses gastrintestinais, caracterizando o manejo sanitário sob fatores condicionantes das formas de controle dessas parasitoses em rebanhos de ovinos da região do Sertão da Paraíba. A pesquisa foi desenvolvida no período de abril a julho de 2012. Foram visitadas propriedades, utilizando-se 465 animais, sendo coletadas individualmente amostras de fezes e sangue durante as visitas. Em cada propriedade, foi aplicado questionário para a coleta de informações acerca de variáveis que atuariam como possíveis fatores de risco. Observou-se que a prevalência das helmintoses gastrintestinais de ovinos na região do Sertão da Paraíba foi de 75,9%. Pelo menos um animal foi positivo para essas helmintoses, em 53 (98,1%) das 54 propriedades avaliadas. A análise de OPG (Ovos Por Gramas de Fezes) demonstrou que 51,8% dos animais apresentaram infecção leve, 27,1% infecção moderada, 9,9% infecção pesada e 11,2% infecção fatal. A utilização de anti-helmínticos ocorreu em 81,5% das propriedades (p <0,05). O fator de risco mais relevante neste estudo foi a área da propriedade, porque delimita a área de pastejo do animal. Propriedades com muitos animais e pouca área de pastejo, que são as mais abundantes no Sertão da Paraíba, tendem a apresentar alta prevalência de helmintoses gastrintestinais, pois os animais estão mais propensos à reinfecção. A região do Sertão da Paraíba apresenta uma elevada prevalência de helmintoses gastrintestinais em ovinos, e a área das propriedades é o fator de risco mais relevante para o desenvolvimento dessas parasitoses.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog’s (MS) medium containing 2.5 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L-1 each and 50 mg L-1 ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and ~40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1 activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.

Genetic characteristics of human acute lymphoblastic leukemia cell line Molt-4 / 中国实验血液学杂志

This study was aimed to investigate the genetic characteristics of human acute lymphoblastic leukemia cell line Molt-4, and evaluate its application in measuring telomere length by Flow-FISH. Molt-4 cell line was cultured in suspension and subcultured regularly. Eight different passages of Molt-4 cells in exponential stage were selected.The growth curves were drawn by cell counting method, meanwhile calculating the population doubling times of cells,DNA ploidies were determined by flow cytometry,karyotypes were analyzed by G-banding and telomere lengths were measured by Southern blot. The results showed that the population doubling time of Molt-4 cell line was (1.315 ± 0.062) d, DNA ploidy index was (2.085 ± 0.0093) , and the telomere length was (32.05 ± 5.27) kb. There were no significant difference among different passages (P = 0.931,0.888 and 0.935 separately). The karyotypes showed that the chromosome numbers of Molt-4 cell line were from 91 to 99 in different metaphases, and the majority of them were hypertetraploid, and stable and recurrent structural abnormalities of chromosomes could be kept. It is concluded that the stable genetic characteristics and the longer telomere length of Molt-4 cell line makes it be a feasible control cells in measurement of telomere length by Flow-FISH.

Estudio de ploidía de células plasmáticas por citometría de flujo en pacientes conmieloma múltiple: primeros casos estudiados en Uruguay / Flow cytometry for plasma cells ploidy analysis in patients with multiple myeloma: first cases studied in Uruguay

Introducción: la historia natural del mieloma múltiple (MM) es heterogénea con sobrevidas que van desde pocas semanas a más de 20 años. El análisis de los factores pronósticos es esencial para establecer una terapéutica adaptada al riesgo. El estudio de la ploidía de las células plasmáticas es un factor que ha demostrado tener un importante valor pronóstico. Objetivo: estandarizar una técnica, no disponible en Uruguay, para determinar la ploidía de las células plasmáticas por citometría de flujo. Material y método: el estudio de ploidía se realizó en médula ósea utilizando para la marcación de las células plasmáticas los anticuerpos monoclonales anti CD38 y CD138. Para el estudio del contenido del ácido desoxirribonucleico (ADN) se utilizó ioduro de propidio. En el análisis se calculó el índice de ADN (cociente entre la moda del pico correspondiente a la cantidad de ADN de las células plasmáticas en fase Go/G1 y la moda del pico Go/G1 de las células normales residuales). Resultados: en este trabajo mostramos la estandarización de la determinación de ploidía por citometría de flujo y los primeros casos analizados en nuestro país. Se estudiaron nueve pacientes con diagnóstico de MM, hallándose dos casos hipoploides (no hiperploide), un caso diploide (no hiperploide) y seis casos hiperploides. Conclusiones: disponemos de una técnica de determinación de ploidía de células plasmáticas que es sencilla, rápida de realizar y con importante valor pronóstico para pacientes portadores de MM.

Introduction: the natural history of multiple myeloma (MM) is heterogeneous, survival rates ranging from a few weeks to over 20 years. Analysis of prognostic factors is essential to decide on a therapy that is adapted to specific risks. Plasma cells ploidy analysis has proved to be a high prognostic value factor.Objective: to standardize a technique, unavailable in Uruguay, consisting in flow cytometry for plasma cells ploidy analysis in order to determine ploidy values in plasma cells.Method: ploidy analysis was performed in the bone marrow, and monoclonal anti-CD38 and CD-138 antibodies were used to mark plasma cells. Propidium iodide was used to study the content of deoxyribonucleic acid (DNA). The DNA was calculated in the analysis (the ratio of the peak mode corresponding to the DNA present in the plasma cells during the Go/G1 phase and the Go/G1 peak mode of residual normal cells). Results:the study presented the standardization of flow cytometry ploidy analysis and the first cases analysed in our country. Nine patients with a diagnosis of MMwere studied, having found two hypoploid cases (non-hyperploid), one diploid case (non-hyperploid),and six hyperploid cases.Conclusions: there is a technique for ploidy determination of plasma cells that is simple, fast to perform and has an important prognostic value for patients with MM.

Introdução: a historia natural do mieloma múltiplo (MM) é heterogênea com sobrevidas que variam de poucas semanas a mais de 20 anos. A análise dos fatores prognósticos é fundamental para estabelecer uma terapêutica adaptada ao risco. O estudo da ploidia das células plasmáticas é um fator que mostrou ter valor prognóstico importante. Objetivo: padronizar uma técnica, não disponível no Uruguai, para determinar a ploidia das células plasmáticas por citometria de fluxo. Material e método: o estudo da ploidia foi reão da determinação da ploidia por citometria de fluxo e os primeiros casos analisados no nosso país. Nove pacientes com diagnóstico de MM foram estudados, sendo encontrados dois casos hipoploides (não hiperploide), um caso diploide (não hiperploide) e seis casos hiperploides. Conclusões: dispomos de uma técnica de determinação de ploidía de células plasmáticas que é simples, rápida de realizar e com valor prognóstico importante para pacientes portadores de MM.

Establishment & characterization of lymphoblastoid cell lines from patients with multiple primary neoplasms in the upper aero-digestive tract & healthy individuals.

Background & objectives: A major drawback for genetic studies as well as long-term genotype-phenotype correlation studies in cancer is lack of representative human cell lines providing a continuous source of basic biomolecules and a system to carry out various experimental investigations. This can be overcome to some extent by establishing lymphoblastoid cell lines (LCLs) by infecting peripheral blood lymphocytes with Epstein Barr virus (EBV) which is known to immortalize human resting B cells in vitro giving rise to actively proliferating B-lymphoblastoid cell lines. The present study involves preparation and characterization of LCLs generated from patients with multiple primary neoplasms (MPN) of upper aero-digestive tract (UADT). Methods: Thirty seven LCLs were established from UADT MPN patients and healthy age, sex and habit matched controls using EBV crude stock. Characterization was done with respect to expression of CD-19 (Pan B-cell marker), CD3 (T cell specific marker), CD56 (NK-cell specific marker), cell morphology, ploidy analysis, genotype and gene expression comparison with the parent lymphocytes. Results: LCLs showed rosette morphology with doubling time of approximately 24 h. Ploidy analysis showed diploid DNA content which was maintained for at least 30 population doublings. When compared with parent lymphocytes there appeared no change at genetic and gene expression level. Interpretation & conclusions: Our results show that lymphoblastoid cell lines are a good surrogate of isolated lymphocytes bearing their close resemblance at genetic and phenotypic level to parent lymphocytes and are a valuable resource for understanding genotype-phenotype interactions.

Production and characterization of a somatic hybrid of Chinese cabbage and cabbage / 生物工程学报

In order to broaden Chinese cabbage gene pool, we conducted interspecific somatic hybridization between Chinese cabbage (Brassica campestris, 2n=20, AA) and Cabbage (B. oleracea, 2n=18, CC). Protoplasts were isolated from 10-day-old cotyledons and hypocotyls of young seedlings, and fused by 40% polyethylene glycol (PEG). Fused cells were cultured in modified K8p liquid medium supplemented with some plant growth regulators. Fusion products were characterized by their morphological, cytological and molecular biological traits. The results showed that, a total of 35 regenerated green plants were obtained from 320 calli, the plant regeneration frequency was 10.94%, and eleven of which were survived in greenhouse. All regenerants were true hybrids as confirmed by randomly amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) analysis. Ploidy levels of hybrid plants were determined by chromosome counting and flow cytometry. The sum of the chromosome number (2n = 38) from the two fusion patents were found in 36.4% of regeneratns; another 36.4% had chromosomes range to 58-60; 27.2% had more chromosomes ranges to 70-76. All regenerated plants produced normal flowers. We investigated the pollen fertility and seed set after self-pollination and backcrossing with the parental species. For hybrids with chromosomes more than 38 it was possible to obtain some seeds when they after self-pollination. Within the group of hybrids with 38 chromosomes, seed set were very variable, only 0.11 seeds per pod by self-pollination, 0.23-0.76 by open-pollination, 0.02-0.04 by backcrossing with Chinese cabbage. Progeny lines obtained by self-pollination had larger leaves and leaf shapes intermediate of the parental species. Pollen fertility was gradually recovered in the first and second progenies. The backcrossing progeny lines, as a whole, exhibited morphologies were similar to Chinese cabbage. Morphological variations were observed among the somatic hybrids and their progenies.

Octaploidy in idiopathic thrombocytopenic purpura.

We report a case of an elderly 68-year-old male who presented in our hospital with chief complaints of petechial rashes and ecchymosis over extremities and bleeding from the oral cavity since 3–4 days prior to hospitalization. He saw a physician before coming to our hospital and received one dose of IV methylprednisolone and oral wysolone. He had come to our hospital for further management. Bone marrow karyotyping was done and chromosomal analysis revealed two cell lines. Eighty percent of the cells analyzed revealed apparently normal male karyotype. However, 20% cells analyzed revealed a total of 184 chromosomes, suggesting octaploidy.

Fas receptor (CD95) & Fas ligand (CD178) expression in patients with tobacco-related intraoral squamous cell carcinoma.

Background & objectives: Fas receptor and Fas Ligand (FasL) system has been implicated in the resistance to apoptosis, insensitivity to chemotherapy and in providing immune privileged status to most of the tumours. However, no reports are available on Fas and FasL expression in patients with tobacco-related oral carcinoma. Therefore, the present study was undertaken to observe Fas and FasL expression and their correlation with clinicopathological features as well as cell cycle parameters. Methods: Immunohistochemistry for Fas, FasL and DNA flow cytometry for cell cycle parameters was successfully done on 41 paraffin embedded tumour and 10 normal samples. The results were evaluated for possible association of Fas and FasL with clinicopathological features and cell cycle parameters. Results: Weak Fas expression was observed on the cell membrane only in 2 of 41 (5%) oral tumours while FasL immunoreactivity was seen in 26 of 41 (63.4%) tumours. In contrast, all ten normal oral tissues exhibited strong cytoplasmic and membrane Fas receptor immunoreactivity but absence of FasL staining. Older patients, greater tumour size and lymph node positivity were found to be associated with high expression of FasL. Significantly higher (P<0.01) expression of FasL was observed in oral tumours with aggressive DNA pattern like aneuploidy and high S-phase fraction. Interpretation & conclusions: Downregulation of Fas receptor and up-regulation of Fas ligand appear to be an important feature of tobacco-related intraoral carcinoma. Association of FasL expression with advanced clinical stage and aggressive DNA pattern suggests that the Fas and FasL system may be used as an important prognostic variable in patients with tobacco-related intraoral squamous cell carcinoma.

Bovine parthenogenotes produced by inhibition of first or second polar bodies emission

Parthenogenetic embryos are an ethically acceptable alternative for the derivation of human embryonic stem cells. In this work, we propose a new strategy to produce bovine parthenogenetic embryos inhibiting the emission of the first polar body during in vitro maturation, and allowing the extrusion of the second polar body during oocyte activation. Cytochalasin B, an inhibitor of actin microfilaments, was employed during in vitro maturation to inhibit first polar body emission or during parthenogenetic activation to block second polar body emission. Only one polar body was inhibited in each strategy in order to keep the diploid chromosome set. In experiment 1, the effect of cytochalasin B on in vitro maturation of bovine oocytes was evaluated. Most oocytes (77%) were arrested at a meiotic stage characterized by the presence of a large internal metaphase plate and absence of polar body. In experiment 2, development of embryos exposed to cytochalasin B during in vitro maturation (CytoB-IVM) or during activation (CytoB-ACT) was compared. Developmental rates did not differ between diploidization strategies, even when three agents were employed to induce activation. Both groups, CytoB-IVM and CytoB-ACT, tended to maintain diploidy. CytoB-IVM parthenogenesis could help to obtain embryos with a higher degree of homology to the oocyte donor.

Comparative study on photosynthetic characteristics of different ploidy Rhodiola sachalinensis germplasm / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the photosynthetic characteristics difference of different ploidy Rhodiola sachalinensis germplasm and provide the scientific basis for their cultivation.</p><p><b>METHOD</b>LI-6400/XT photosynthesis system was used to measure leaf light response curve and CO2 response curve of diploid and autotetraploid. Biomass, leaf area, stomatal characteristics and chlorophyll content differences were compared in the study.</p><p><b>RESULT</b>Stomata of the two germplasms were open during daytime obviously, and stomata conductance responded to the changes of light intensity and CO2 concentration which was not consistent with the characteristics of CAM (crassulacean acid metabolism) plants. Light compensation point of autotetraploid was significantly lower than that of the diploid, and light saturation points of both germplam were close, and their light saturation points were near 500 micromol x m(-2) x s(-1). Quantum efficiency of autotetraploid was significantly higher than the diploid, and the net photosynthetic rate of autotetraploid significantly higher than the diploid when light intensity was higher than 500 micromol x m(-2) x s(-1). Stomata conductance, transpiration rate of autotetraploid was also significantly higher than that of diploid. Biomass, leaf area, stomata diameter and chlorophyll content of autotetraploid were much higher than that of diploid, while the stomata density of autotetraploid was less than diploid.</p><p><b>CONCLUSION</b>The results above provide scientific basis for the cultivation of different ploidy Rh. sachalinensi germplasm.</p>