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Objective: To explore the genetic and clinical characteristics of near-tetraploidy/tetraploidy karyotype (NT/T) in patients with myelodysplastic syndrome (MDS)., Methods: Cytogenetic findings of 1576 inpatients with primary MDS were retrospective analyzed, among which 9 were diagnosed with NT/T. Clinical data including gender, age, morphology, genetic feature and prognosis were analyzed., Results: The prevalence of MDS patients with NT/T (NT/T-MDS) among all cases was 0.57%. Karyotyping analysis suggested that eight MDS patients had sole NT/T, while the remainder one had a complex karyotype. In addition to the typical morphology of MDS, NT/T-MDS had unique morphology including huge blast, double-nuclear cell and irregular nuclear membrane. One NT/T-MDS patient gave up therapy, and the remaining eight underwent the first course of treatment, albeit with poor prognosis. Only one patient had complete remission, one had partial remission, three had no remission; and three had converted to acute myeloid leukemia., Conclusion: NT/T-MDS is rare and has unique morphology. Generally, NT/T-MDS patients have poor prognosis. However, NT/T cannot be simply classified as high-risk group, but with consideration whether they have affected particular chromosomal structures as well as other clinical data.

Parthenogenetic embryonic stem cells (pESCs) derived from bi-maternal genomes do not have competency of tetraploid complementation, due to lacking of paternal imprinting genes. To make pESCs possess fully development potentials and similar pluripotency to zygote-derived ESCs, we knocked out one allelic gene of the two essential maternal imprinting genes (H19 and IG) in their differentially methylated regions (DMR) via CRISPR/Cas9 system and obtained double knock out (DKO) pESCs. Maternal pESCs had similar morphology, expression levels of pluripotent makers and in vitro neural differentiation potentials to zygotes-derived ESCs. Besides that, DKO pESCs could contribute to full-term fetuses through tetraploid complementation, proving that they held fully development potentials. Derivation of DKO pESCs provided a type of major histocompatibility complex (MHC) matched pluripotent stem cells, which would benefit research in regenerative medicine.

Objective: To analyze a fetus with abnormal cardiac ultrasound by using various techniques and explore its genotype-phenotype correlation., Methods: Lymphocytes derived from umbilical cord blood sample were subjected to G-banding analysis. Short tandem repeats quantitative fluorescence PCR (STR-QF-PCR) was used for analysis of fetal DNA as an auxiliary test. Low-coverage whole genome sequencing (WGS) was used to detect chromosomal deletion/duplication which exceeded 100 kb in size., Results: The karyotype of the fetus was 47,XN,+mar. As detected by STR-QF-PCR, the copy number of GATA178F11 locus on chromosome 18 was 4, and the duplicated fragment was derived from the mother. WGS suggested that the fetus to be 46,XN,dup(18p11.21p11.32).seq [GRCh37/hg19](10 001-15 378 887)× 4, with the duplicated fragment spanning approximately 15.38 Mb., Conclusion: The cardiac malformation of the fetus may be attributed to the partial duplication of chromosome 18p. Combined cytogenetic and molecular methods can facilitate prenatal detection of genetic abnormalities.

Objective: To explore the genetic and clinical features of patients with acute myeloid leukemia (AML) and near-tetraploidy/tetraploidy (NT/T) karyotype., Methods: Cytogenetic findings of 1836 cases of primary AML were retrospectively analyzed. Karyotypes of the identified cases were confirmed by fluorescence in situ hybridization (FISH). Clinical data including gender, age, morphology, immunophenotype, genetics, and prognosis were reviewed., Results: Nine male and two female patients with NT/T were identified with a median age of 63 years. Microscopically, the patients were characterized by large blasts and irregular nuclear contours. All patients expressed CD34, and nine of them expressed HLA-DR. Ten patients had complete remission during the first course of treatment. One patient showed primary drug resistance., Conclusion: NT/T AML primarily occurs in elder males and has a characteristic morphology and genetics. The prognosis is better than AML patients with complex karyotypes.

Objective: To diagnose chromosomal abnormalities in amniotic fluid cells by combining karyotyping and single nucleotide polymorphism array (SNP-array) analysis, and to explore the application of SNP-array in routine clinical practice., Methods: Conventional G banding was used to karyotype a fetal amniotic fluid sample and the corresponding peripheral blood samples from the parents, followed by SNP-array analysis of the fetal genomic DNA from the amniotic fluid., Results: The karyotype of the amniocytes was 47, XX, +mar. The marker chromosome was further identified as psu idic (22) (q11.2) by SNP-array analysis, revealing tetraploidy of a 1.7 Mb fragment in 22q11.1-q11.2 interval that involves the critical region for Cat eye syndrome., Conclusion: A rare chromosomal abnormality was identified by combining conventional G banding and SNP-array. The high resolution SNP-array could provide more detailed information for determining the origin of chromosomal abnormalities.

In order to investigate the epigenetic variations between diploid and autotetraploid of Platycodon grandiflorus. The diploid buds of P. grandiflorus were soaked in the mixture of different concentration colchicines and 0.002 g•mL ⁻¹ dimethyl sulphoxide (DMSO)．The identification of autotetraploid plants were based on morphological characteristics, chromosome number and flow cytometry. And then the level and pattern of DNA methylation explored by using the technology of methylation sensitive amplified polymorphism (MSAP)．The result demonstrated that the buds soaked in 0.2% colchicines and 0.002 g•mL ⁻¹ DMSO solution for 12 h was ideal conditions to induce autotetraploid of P. grandiflorus, with induction rate of 32.0%.The diploid and tetraploid plants existed distinctly differences in morphological indexes.Totally,1 586 bands were amplified by 20 pairs of selective primers, of which 764 and 822 bands were detected in diploid and autotetraploid respectively. The total methylation ratio,full methylation ratio and hemimethylated ratio were 91.25%,61.25% and 30.65% in diploid of P. grandiflorus,respectively.However,the total methylation ratio,full methylation ratio and hemimethylated ratio of autotetraploid of P. grandiflorus were 86.13%,54.38% and 31.75%, respectively. Compared with diploid, the genomic DNA total methylate ratio and full methylation ratio of autotetration plants decreased by 6.02% and 7.14%.But the hemimethylated ratio of autotetraploid was higher than that of diploid, which more than 1.6%. All this results indicated that DNA methylation patterns have adjusted during the polyploidy process．., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

In order to investigate the genetic basis of morphological variation of tetraploid plantlets of Atractylodes macrocephala, diploid plantlets were taken as experimental material, sterile filtration colchicine was used to soak 0.5-1.0 cm long buds. The difference between morphology and stomatal of diploid and tetraploid of A. macrocephala was compared, and genome polymorphism was explored by AFLP. The results showed that the buds dipped in 0.1% colchicine solution for 36 h was optimal conditions to induce tetraploid of A. macrocephala with induction rate of 32.0%. Morphological indexes such as leaf area index, leaf length and width, the density of stomas and the number of chloroplast of tetraploid were distinctly different from diploid. Four hundred and fifty-one bands ranging with 80-500 bp were amplified with 24 pairs of primers, the rate of polymorphism was 32.59%. These amplification sites of diploid were different from tetraploid of A. macrocephala, and the differences in morphology of them were reflected in the DNA polymorphism.

Simple sequence repeats (SSRs)are a class of repetitive DNA sequences, which are commonly used for genome analysis. Comparison of the homologous SSRs among different genomes is helpful to understand the evolutionary process in relative species. In this study, SSR scanning was performed to investigate their distribution and length variation among the genomes of G. raimondii (D₅), G. arboretum (A₂) and G. hirsutum (AD₁). The results demonstrated that the distribution of SSRs in A genome was very similar with that in D genome, while the length variation of homologous SSRs between A and AD genome was more conserved than that between D and AD genome. Compared with SSRs in AD genome, the number of SSRs with longer motif length in A genome was about five times of those with shorter motif length, while it was about three times in D genome. This implied that the length variation rates of homologous SSRs between diploid cotton and tetraploid cotton were different during the parallel evolution due to the subgenome fusion, and the motif length of most SSRs in tetraoploid genome tended to become shorter than homologous SSRs in diploid genome during the process of evolution. This study comprehensively compared the SSRs in three cotton genomes and revealed the significant difference among them, providing a foundation for further evolutionary study of Gossypium genome.

Objective: To provide technical support for industrialization promotion of tetraploid of Dioscorea zingiberensis, the manufacturing method for synthetic seeds of tetraploid of Dioscorea zingiberensis was established and the correlated influential factors were studied., Methods: By taking embryogenic calluses of tetraploid of Dioscorea zingiberensis as propagation materials, the influential factors such as components of artificial endosperm, seed coats,storage conditions and germination materials on germination and seedling of the synthetic seeds were evaluated., Results: When 4% alginate +2% CaCl2 + 2% chitosan was served as seed coat materials, and 1/2 MS +0. 2 mg/L BA +0. 5 mg/L NAA + 0. 1 mg/L penicillin + 0. 3% carhendazim powder + 0. 2% sodium benzoate + 1. 0% sucrose + 0. 5% activated carbon + 1. 0% tapioca starch was served as endosperm, the synthetic seeds had high germination rate and seedling rate. After storing at 4 °C for 20 d, the germination rate and seedling rate of synthetic seeds was 76. 7% and 71. 7%, respectively., Conclusion: Manufacturing technology of synthetic seeds of tetraploid of Dioscorea zingiberensis with embryogenic calluses as propagation materials has production prospects.

Objective: To identify tetraploid Isatis indigotica strains through morphology and flow cytometry., Methods: The tissue culture seedlings of tetraploid Isatis indigotica were root-tip squashed and chromosome counted before rooted climatized and transplanted in field. The plants in field were taken as experimental materials. Macroscopic observation was applied to identify by form and structure; Free-hand section was used to observe the length, width and density of stomas; And flow cytometry was applied to identify the ploidy., Results: Compared with diploid plants, tetraploid plants had obvious changes in form and structure. The stomas from the tetraploid were notably longer, and the number of guard cells in chloroplasts was remarkably larger. The experiment materials were proved to be tetraploid by flow cytometry., Conclusion: The materials are tetraploid plants. Macroscopic observation, the length of stoma and the number of guard cells in chloroplasts can be taken as aided identification for ploidy of mutagenesis materials. Meanwhile, flow cytometry can be applied to identify the ploidy of Isatis indigotica.

Objective: To study the genetic stability of autotetraploid plant of Dioscorea zingiberensis., Methods: The chromosome of root-tip was determined by photomicroscope, and the agronomic characters were observed in the period of stable growth. The protein content was determined and the experiment of protein polyacrylamide gel electrophoresis was carried out. Furthemore, the diosgenin content was determined and compared., Results: The chromosome number of autotetraploid plantlet was 2n = 4x = 40. The agronomic characters showed typical autotetraploid characteristics. The contents of diosgenin and protein of autotetraploid were higher than that of the diploid. The protein electrophoresis bands of all the lines were similar., Conclusion: The experiment confirmed that the autotetraploid plant of Dioscorea zingiberensis, which was artificially induced, had good genetic stability. It lays the foundation for the polyploid breeding to develop superior varieties of Dioscorea zingiberensis.

Objective: To establish and optimize the rapid propagation system of Polygonum multiflorum, as well as explore method for induction and identification of autotetraploid., Method: Propagation medium was optimized by orthogonal test. The buds were immersed in colchicine solution with different concentrations for different time to select induction conditions for autotetraploid of P. multiflorum., Result: The most appropriate propagation medium was MS medium supplemented with 1.0 mg x L(-1) 6-BA, 0.3 mg x L(-1) NAA, and 0.4 mg x L(-1) PP333. That the buds were soaked in 0.2% colchicine solution for 30 h, or soaked in 0.3% colchicine solution for 18 h, was optimal condition to induce autopolyploid of P. multiflorum with induction rate as high as 16.7%., Conclusion: Rapid propagation of P. multiflorum could be achieved by tissue culture. Furthermore, colchicine was an effective inducer of polyploidy, and 25 tetraploid lines were obtained through chromosome identification. The experiment laid a foundation for the wild resource conservation, superior varieties breeding of P. multiflorum.

Objective: To provide a reference evidence for choosing salt-resistent and drought-resistent varieties of Isatis indigotica Fort.., Methods: The tissue culture seedlings of different Isatis indigotica tetraploids were used as plant materials. The diploid was used as CK, which were cultured for 30 days. 2 g/L NaCl and 15% PEG-6000 were used as stress treatment respectively for 12 hours, then poured out the solution, the materials were continuously cultured for 7 days. The contents of indirubin and indigotin were determined by HPLC., Results: The contents of indigotin and indirubin of DB3, DB5 and DB12 with PEG treatment and DB2, DB3 and DB12 with NaCl treatment were increased., Conclusion: DB3 and DB12 can be further studied as reproduction materials.

Objective: To study the effect of gas-turbine green discoloring and drying processing method on the quality of various Lonicerae Japonicae Flos herbs., Method: DIKMA DiamonsilTM-C18 column (4.6 mm x 250 mm, 5 microm) was adopted using HPLC Waters 1525 and eluted with acetonitrile and 0.1% phosphate acid as the mobile phase. The flow rate was 1.0 mL x min(-1) , the column temperature was 25 degrees C the detection wavelength was 355 nm., Result: After being processed by the gas-turbine green discoloring and drying method, tetraploid Lonicerae Japonicae Flos showed a green color. The contents of chlorogenic acid and galuteolin were 5.31% and 0.105% , both significantly higher by 18.0% and 32.1% than those of diploid Lonicerae Japonicae Flos processed by the same method. The content of chlorogenic acid in tetraploid Lonicerae Japonicae Flos processed the gas-turbine green discoloring and drying method were also remarkably higher than that of tetraploid and diploid Lonicerae Japonicae Flos processed by traditional processing method of natural drying., Conclusion: The gas-turbine green discoloring and drying processing method is a new-type drying method suitable for tetraploid Lonicerae Japonicae Flos. Under the condition of gas-turbine green discoloring and drying processing, tetraploid Lonicerae Japonicae Flos shows much higher quality than Lonicerae Japonicae Flos, suggesting that it is a good variety worth popularizing and applying.

Protein kinase C (PKC) is a critical molecule in cellular signal transduction in mammals. It is involved in many biological processes in embryonic development, including nuclear remodeling, cell cycle adjustment and cellular polarity regulation. The present study aimed to observe the location of PKCα, an important isozyme of PKC, in fertilized, parthenogenetic and tetraploid preimplantation embryos, and compare the expression of PKCα during embryonic compaction in Kunming mice. The location of PKCα was detected by immunochemistry and laser confocal microscopy. Western blot was performed to quantify PKCα expression during embryonic compaction in the three kinds of embryos. In the experiment, fertilized embryos were flushed from oviduct or uterus at 45, 52, 69, 76 and 93 h after injection of human chorionic gonadotrophin (hCG); parthenogenetic embryos were collected by SrCl2 activation of oocytes for 6 h; and tetraploid embryos were produced by electrofusion of 2-cell embryos. Embryos were fixed at different developmental stages for immunofluorescent staining. 8-cell/4-cell embryos and morula were lysed for Western blot. The results showed that PKCα had similar location pattern in different embryos. It was distributed mainly in the nuclear aggregating around chromatin at different developmental stages. However, PKCα expressed strongly in the interphase than in mitotic blastomere. Before embryonic compaction, PKCα was localized at the blastomere boundary. At late blastocyst stage of fertilized embryos, PKCα was localized only in the polar trophoblast, but not in other trophoblast. At late stage of pathenogenetic blastocyst, there was no clear PKCα signal in the polar trophoblast. Tetraploid embryos had larger blastomere than other embryos and compacted after 4-cell stage, but not after 8-cell stage. Meanwhile, there was PKCα signal at the blastomere boundary at 4-cell stage. Our results showed that the expression of PKCα lasted through all the preimplantation stage. Although there were different expression levels among different stages, the expression increased around embryonic compaction. Quantification of expression of PKCα by Western blot demonstrated that the expression increased after compaction, indicating that the compaction was possibly dependent on the relocation of PKCα. Moreover, it was shown that the second relocation of PKCα occurred during the blastocyst formation. PKCα had different expression patterns in the three kinds of preimplantation embryos. However, the effects of PKCα on embryonic development started in early stage. There must be a necessary connection between PKCα relocation and cell adhesion starting at embryonic compaction.