超高速细胞分选平台结合cDNA microarray技术筛查宫颈癌细胞潜在分子标志物

目的:通过超高速细胞分选平台结合cDNA microarray技术,筛查宫颈癌细胞可能潜在的分子标志物。方法:采用MoFlo XDP型超高速细胞分选平台纯化细胞膜表面表达CD38和不表达CD38的宫颈癌细胞,利用RNAlater技术得到cDNA microarray实验所需RNA,然后进行基因芯片分析。结果:利用MoFlo XDP型超高速细胞分选平台可以获得纯度为99.0%以上的CD38阳性表达宫颈癌细胞。结论:cDNA microarray分析发现了RORA、PLIN4、AUTS2、IFITM1等宫颈癌细胞潜在分子标志物,为宫颈癌研究提供了新的技术方法。

Correlation between Reasons for Prescription and Karyotype Results in Patients Referred for Suspected Chromosomal Abnormalities

Karyotype prescription is based on clinical signs (or reasons for karyotype prescription) which are phenotypic manifestations associated with chromosomal abnormalities. The aim of this study was to establish a correspondence between karyotype indications and their results in patients. This was a retrospective study that was carried out in the Histology-Embryology-Cytogenetics laboratory of the University Hospital of Cocody-Abidjan from 2014 to 2019. 58 patient files were identified and included the indication or reason for prescribing a constitutional karyotype and the biological result obtained. An individual data sheet was used to collect the data. 17 reasons for prescription were identified and divided into 2 groups. Sexual ambiguity was the most frequent reason (29.3%). The first group (G1) represented the 10 reasons for which the karyotype results were normal. The second group (G2) corresponded of the 7 motives with normal or abnormal karyotype results. Several anomalies were listed according to these reasons: inversions, mosaics (anomalies of number and structure) and trisomy 21. The last was the most frequent chromosomal anomaly (69.24%). It was found in several reasons for karyotype prescription: malformations, neurological disorders, suspected trisomy and cardiac pathology. Several factors could explain these results, among which are the limits of the karyotype and the non-genetic causes that can induce these abnormal phenotypes. Complementary examinations to the karyotype are molecular cytogenetic techniques, notably fluorescence in situ hybridization (FISH) and array comparative genomic hybridization (Array-CGH).

Prospects of DNA microarray application in management of chronic obstructive pulmonary disease:A systematic review

Chronic obstructive pulmonary disease(COPD)is incurable chronic disease which kills 3.3 million each year worldwide.Number of global cases of COPD is steadily rising alongside with life expectancy,disproportionally hitting middle-income countries like Russia and China,in such conditions,new approaches to the COPD management are desperately needed.DNA microarray technology is a powerful genomic tool that has the potential to uncover underlying COPD biological alteration and brings up revolutionized treatment option to clinicians.We executed systematic review studies of studies published in last 10 years regarding DNA microarray application in COPD management,with complacence to PRISMA criteria and using PubMed and Medline data bases as data source.Out of 920 identified papers,39 were included in the final analysis.We concluded that Genome-wide expression profiling using DNA microarray technology has great potential in enhancing COPD management.Current studied proofed this method is reliable and possesses many potential applications such as individual at risk of COPD development recognition,early diagnosis of disease,COPD phenotype identification,exacerbation prediction,personalized treatment optioning and prospect of oncogenesis evaluation in patients with COPD.Despite all the proofed benefits of this technology,researchers are still in the early stage of exploring it’s potential.Therefore,large clinical trials are still needed to set up standard for DNA microarray techniques usage implementation in COPD management guidelines,subsequently giving opportunity to clinicians for controlling or even eliminating COPD entirely.

Studies on the Karyotype of Two Species in Orius Wolff (Hemiptera: Heteroptera: Anthocoridae)

The karyotype of two Chinese species in Orius were studied in male germ cells prepared on air dried slides stained with Giemsa. It is reported that two species have 24 chromosomes in diploid and X Y sex chromosomal mechanism. But the beha vior of chromosomes during meiosis between two species is different. Which show in the arrangment and location of the X chromosome and autosome at the late diakinesis as well as metaphase stage respectively and so on. These characteristics may be used in ta xonomy among species of Orius .

Karyotype Study on Scilla scilloides (Lindl.) Druce in Yantai

[Objective] The aim of this study was to investigate the chromosome number and the karyotype of Scilla scilloides in Yantai.[Method]Root tips of Scilla scilloides were pretreated by 8-hydroxyquinoline,then fixed,dissociated and stained for slice production.The chromosome number was analyzed by microscopic examination,and then cells with good chromosomal morphology and dispersal chromosome were studied by microscopic photos.[Result]The somatic chromosome number of Scilla scilloides in Yantai was 2n=16+1Bs,while the karyotype formula was K(2n)=2x=16+1Bs=6m+4sm(2SAT)+4st+2t+1Bs and the karyotype classification was "3B" type.[Conclusion]Karyotype comparison,division of cell type and evolution of Scilla scilloides in Yantai are discussed,which provides basis for cytogenetics,evolutionary genetics,modern taxonomy and genetic breeding.

Studies on the Karyotype of Three Species in the Genus Orius Wolff (Hemiptera:Anthocoridae)

Karyotype of 3 species of the genus Orius Wolff were firstly studied using the Giemsa staining and squashing slides method in male germ cell. It is reported that number of chromosome and system of sex chromosome of haploid chromosome complement of these species is n=11A＋X （Y） in the first meiotic stage. However, the behavior of chromosomes among the species are different distinctly, thus cytotaxonomic character can be adopted to the identification among the three species.

Karyotype Analysis of Lycium ruthenicum Murr.

Karyotype analysis of Lycium ruthenicum Murr. was carried out in this study. The results showed that the chromosome number was 2n=2x=24; the arm index was 48; the ratio of the longest chromosome to the shortest one was 1.31; the proportions of chromosomes with arm ratio higher than 2 was 0.08; the asymmetry index was 57.02; the karyotype type was 2A; and the karyotype formula was 2n=-24=20m＋4sm.

Morphological Difference and Karyotype of Pelteobagrus fulvidraco in Dongting Lake Water System

[Objective] The research aimed to study the morphological characteristics and karyotype of Pelteobagrus fulvidraco in Dongting Lake water system.[Method] By using the conventional biological morphometry,PHA and colchicine injection method in vivo,the morphological characteristics and karyotype of P.fulvidraco in Yuanshui River and Lishui River of Dongting Lake were analyzed.[Result] In three ratio traits including standard length/head length,standard length/caudal peduncle depth,head length/snout length,P.fulvidraco of Yuanshui River and Lishui River had significant differences（P0.05）.However,the number and karyotype of their chromosomes were same.The chromosome number was 2n = 52,and the karyotype formula was 20M＋12SM＋10ST＋10T.The number of chromosome arm was 84.[Conclusion] The research result had certain theoretical guidance significance for the protection and utilization of wild P.resource in Dongting Lake water system.

The Karyotype of Chi-lin Fish(Varicorhinus macrolepis) from Taishan Mountain

[Objective] The paper was to study the karyotype of Chi-lin fish （Varicorhi- nus macrolepis） from Taishan mountain. [Method] The chromosome number and karyotype of an endemic Chi-lin fish were studied using the head kidney cells. [Re- sult] The diploid chromosome number （2n） of Chi-lin fish was 50, and the karyotype of V. macrolepis was 2n=50,16m＋14sm＋20t, NF =80. No sex-chromosome was found in Chi-lin fish. [Conclusion] A detailed karyotype of this endemic cyprinid fish of Taishan Mountain was established for the first time in this study.

Chromosomal Karyotypes of Fragaria Plants

[Objective]This study was to analyze the karyotypes of wild diploid strawberry species,so as to provide basis for revealing the ori- gins,evolvement and genetic breeding of strawberry. [Method] Four diploid strawberry species including Fragaria gracilis A. Los,Fragaria nilgerrensis Schidl,Fragaria pentaphylla A. Los. and Fragaria vesca L. were used as experimental materials to analyze their chromosomal kary- otypes. [Result]The karyotype formula for strawberry species tested was as follows： 2n =2x =14 =10m ＋2sm ＋2m＊ for F. gracilis A. Los, belonging to type 1B; 2n =2x =14 =14m for F. nilgerrensis Schidl,belonging to type 1A; 2n =2x =14 =12m ＋2sm for F. pentaphylla A. Loz. , belonging to type 1A; 2n =2x =14 =14m for F. vesca L. ,belonging to type 1A. Karyotype analysis showed that karyotypic asymmetry degree of the four species was F. gracilis A. Los. F. pentaphylla A. Loz. F. vesca L. F. nilgerrensis Schidl. [Conclusion] The evolution order of these four species was probably F. gracilis A. Los. ,F. pentaphylla A. Loz. ,F. vesca L. ,F. nilgerrensis Schidl.

Research on the Karyotype and Evolution of Drosophila melanogaster Species Group

Mitotic metaphase chromosomes of 34 species of Drosophila melanogaster species group were examined. Certain new karyotypes were described for the first time, and their evolutionary and interspecific genetic relationships among 8 subgroups of D. melanogaster species group were analyzed systematically. The results were as follows. The basic karyotype of elegans subgroup was type A. The karyotypes of eugracilis subgroup, melanogaster subgroup, and ficusphila subgroup were all type C. The karyotypes of takahashii subgroup and suzukii subgroup were both type C and type D. The montium subgroup had six kinds of karyotypes types B, C, C＇, D, D＇, and E. The ananassae subgroup had three kinds of karyotypes： types F, G, and H. Thus, the melanogaster species group was classified into five pedigrees based on the diversity of these karyotypes： 1） elegans; 2） eugracilis-melanogasterficusphila; 3） takkahashii-suzukii; 4） montium; 5） ananassae. The above-mentioned results in karyotypic evolution were consistent with those of DNA sequence analysis reported by Yang except for the elegans subgroup and this subgroup was considered as the ancestral subgroup. Karyotype analysis of the same drosophila from different isofemale lines indicated that the same Drosophila from different places showed karyotypic variation which might be due to different geographical environment and evolutionary degree or interaction between the two factors.

Expression and Significance of Survivin mRNA in Lung Cancer of Different Progression Stages by FISH and Tissue Microarray Technology*

Objective： To investigate the expression of Survivin mRNA in lung cancer progression tissue microarray by FISH （fluorescence in situ hybridization） method and determine its role and significance in lung cancer genesis and progress. Methods： The expression of Survivin mRNA was detected by FISH method and tissue microarray technology. 89 cases of primary lung cancer, 12 cases of lymph node metastasis of lung cancer, 12 cases of precancerous lesion and 10 cases of normal lung tissue were examined. Results： 69.7% of primary lung cancer express Survivin mRNA; the positive ratio of primary lung cancer and precancerous lesion were both significantly higher than that of normal lung tissue （P〈0.05）; the expression of Survivin mRNA was related to the differentiation degree, lymph node metastasis and clinical stages （P〈0.05）. Conclusion： FISH has good sensitivity and stability. Tissue microarray technology has many advantages, such as high efficiency, high throughput, etc; it may have good prospect in pathology. Survivin mRNA was highly expressed in lung cancer and precancerous lesion; it was related to the progress and malignant behavior; it may play a promotion role in lung cancer genesis and progress and offer basis to early diagnosis, prognosis estimate and treatment.

Expression of COX-2 in Different Subtypes of Gastric Intestinal Metaplasia and Gastric Carcinoma by Tissue Microarray

Objective: To study the expression of cyclooxygenase-2 (COX-2) protein in different subtypes of intestinal metaplasia (IM) and gastric carcinoma, evaluate the possibility of COX-2 forecasting the risk of malignant potential of IM, and the relationship between COX-2 expression and gastric carcinogenesis. Methods: Forty cases of chronic atrophic gastritis (CAG) with IM, 40 cases of gastric carcinoma and corresponding paracancerous tissues were selected to construct a tissue microarray. High iron diamine/alcian blue (HID/AB) staining and Hematoxylin and Eosin (HE) staining was used to classify IM and gastric carcinoma, and the expression of COX-2 protein detected in different subtypes of IM and gastric cancer by using immunohistochemistry. Results: The positive expression rate of COX-2 was 45.65%, 59.38% and 77.27% in IM foci in CAG, IM foci in paracancerous tissues, and intestinal-type gastric carcinoma, respectively, significantly higher than in diffuse-type gastric cancer (16.67%)(P<0.05, 0.005 and 0.005, respectively), and the expression intensity of COX-2 protein showed a increased tendency gradually in the sequence of IM foci in CAG→IM foci in paracancerous tissues→intestinal-type gastric carcinoma (P<0.005). The positive expression rate of COX-2 protein in type Ⅲ IM was significantly higher than in type Ⅰ and type Ⅱ IM (P<0.005 and 0.05, respectively), and the expression intensity also showed a increased tendency gradually from type Ⅰ to type Ⅲ IM (P<0.005). Conclusion: The expression level of COX-2 was increased gradually along with the increase of the risk of malignancy of IM, and its expression level may be a useful index to forecast the risk of malignant potential of IM. COX-2 expression was associated with intestinal-type gastric carcinoma, but it might also have some role in the carcinogenesis of diffuse-type gastric carcinoma.

Comparative study of the chromosome karyotypes of two species of grasshoppers in Megaulacobothrus Caudell (Orthoptera: Acridoidea)

The chromosome karyotypes of Chorthippus （Megaulacobothrus） aethalinus （Zubovsky） and Ch. （M.） chinensis Tarbinsky were compared. Results show that the chromosome numbers of the two species fit 2n （♂） = 17 = 16 ＋ XO, in which three pairs of autosomal and sex chromosomes were terminal chromosomes, and the other five pairs of autosomes were metacentric chromosomes. These are diagnosis characters for Chorthippus. However, these two species can be distinguished by the different chromosome formula （CF） and the relative length （RL）. The CF in Ch. （M.） aethalinus is K （2n, ♂） = 6m ＋ 11t = 6L ＋ 6M ＋ 4S ＋ XO, whereas that of Ch. （M.） chinensis is K （2n, ♂） = 6m ＋ 11t = 6L ＋ 8M ＋ 2S ＋ XO. In addition, we also found that the relative lengths of sex chromosomes in the two species were different from each other. The sex-chromosome of Ch. （M.） aethalinus is located at the fifth position and its relative length is 8.33% whereas that of Ch. （M.） chinensis is at the eighth position and its relative length is 5.53%. These results show that significantly different genetic features exist for the two species of Chorthippus.

A new species in the genus Hedotettix Bolivar(Orthoptera: Tetrigidae), including chromosome karyotype, from the western Yunnan Province in China

A new species, Hedotettix nujiangensis Zheng sp. nov., is described. The chromosome complement of H. nujiangensis consists of 2n （♂） = 13. Sex determination is XO. All chromosomes are telocentric （T） and the sex chromosome is the fourth element in size. Type specimens are deposited at Southwest Forestry University.

Comparative study of the karyotypes of two species of grasshoppers in the genera Chorthippus Fieber and Euchorthippus Tarbinsky(Orthoptera: Arcypteridae)

The chromosome karyotypes of Chorthippus albomarginatus （De Geer） which represents the genus Chorthippus Fieber and Euchorthippus unicolor （Ikonnikov） which represents the genus Euchorthippus Tarbinsky were compared. The chromosome numbers for these two species fit 2n （♂） = 17 = 16 ＋ XO, in which five pairs of autosomal and sex chromosomes are terminal chromosomes, and the other three pairs of autosomes are metacentric chromosomes. This similarity between the two genera indicates that they have a close phylogenetic relationship. However, these two species can be distinguished by their different chromosome formula （CF） and the relative sex chromosome length （RL）. The CF in C. albomarginatus is K （2n,♂） = 6m ＋ 11t = 8L ＋ 4M ＋ 4S ＋ XO, whereas that ofE. unicolor is K （2n, ♂） = 6m ＋ 11t = 8L ＋ 6M ＋ 2S ＋ XO. The relative lengths of sex chromosomes in the two species are different. The sex-chromosome of C. albomarginatus is located at the ninth position and its relative length is 2.96% whereas that ofE. unicolor is at the eighth position and its relative length is 4.26%. These results show that significantly different genetic features exist for the two genera.

Chromosomes and Karyotypes of Three Seagrass Species of Hydrocharitaceae

[Objective] This study aimed to investigate the chromosomes and karyotypes of three seagrass species of hydrocharitaceae-Enhalus acoroides, Thalassia hemprichii and Halophila minor collected from Li’an Lagoon, Hainan Province, China. [Method] The root-tips of E. acoroides, T. hemprichii and the axillary buds of H. minor were selected as the materials in this study. The conventional crushing method was used to prepare the slice specimens of chromosomes, and the karyotypes of the three species were analyzed. [Result] The chromosome numbers of E. acoroides, T. hemprichii and H. minor were 2n=18, 18, 28, respectively. The karyotype formulas were K （2n）=18= 12m＋6sm, K（2n）=18=12m＋6sm and K（2n）=28=16m＋8sm＋4st, respectively. According to the standard of Stebbins, the karyotypes of E. acoroides and T. hemprichii were 2A, and that of H. minor was 2B. There was no B chromosome and satellite in these three species. [Conclusion] According to the comparison on the karyotypes of this three species, E. acoroides and T. hemprichii are similar in karyotypes to each other, indicating that there is close, inter-relationship between E. acoroides and T. hemprichii.

A COMPARATIVE STUDY ON KARYOTYPES AND CHROMOSOME BANDING PATTERN OF THREE ODOR FROGS OF Rana

The chromosomes of three odor frogs, Luetuosae- group of Rana were analyzed by conventional, as well as Obanding and silver staining techniques. The three species here examined all had 26 chromosome karyotypes encompassing 5 large and 8 small homologous pairs. Differences among them were found in gross shapes of chromosomes and positions of secondary constrictions. Generally speaking, karyotypes of R. andersonii and R. grahami resembled each other, while the karyotype of R. tiannanensis differed from the former two in several respects.Analyses of karyotypes by C-banding technique indicated that, centromeric areas of every chromosome and interstitial parts of some chromosomes of each species were hetero-chromatinized, and differences of distribution of heterochromatin were found among species. In early metaphase plates of R. andersonii, much more heterochromatinized areas were observed, and when it reached late metaphase, the numbers of heterochromatin sections in each chromosome pair reduced to a limited level, e. g. centromeres and a few interstitial parts.The active nucleolar organizer regions (NORs) were localized in long arms of pair No. 10, in connection with secondary constrictions for R. andersonii and R. grahami as in other odor frogs previously reported, but in the long arm of pair No. 6 for R. tiannanensis.The cytogenetic and taxonomic implications of the findings were discussed based on comparisons with each other, and with published literature.

Identification of p63 expression in human lung cancer: analysis by complementary DNA and tissue microarray

Objective: To evaluate p63 expression at mRNA transcripts and protein levels in lung squamous cell cancer (SCC), adenocarcinoma, large cell lung cancer (LCLC) and small cell lung cancer (SCLC) and their matched metastatic tumors. The association between p63 expression and p63 locus at chromosomal 3q27 q29 was also investigated. Methods: p63 mRNA expression levels in a large series of lung cancers including SCC, adenocarcinoma, LCLC, SCLC and their matched metastatic tumors were analyzed by cDNA microarray technology. A tissue microarray from 150 primary lung cancer specimens was constructed and used for immunohistochemical detection of p63 protein expression. Chromosomal imbalances at the p63 locus in 70 primary lung cancers samples were studied by comparative genomic hybridization (CGH) technology. Results: mRNA levels were 10 fold in SCC compared to LCLC, SCLC, and adenocarcinoma. Interestingly, the mRNA expression of p63 in metastatic carcinomas was significantly higher than that in their matched primary tumors ( P <0 001). Immunohistochemistry demonstrated that p63 expression was 94.64% in SCC but only 1 79% in lung adenocarcinoma and 2 of 4 LCLC were positive staining. All the results in of SCLC were negative. There was a statistically significant difference for p63 positivity between pT1 tumors and those of higher stage ( P =0 035). The CGH results indicated that p63 locus at chromosomal 3q27 q29 was overrepresented in SCC. p63 immunopositivity correlated significantly with pronounced gains of the p63 locus at chromosomal 3q27 q29 (P=0.0001), indicating that strong expression of p63 in lung SCC correlated with increased gene amplification. Conclusion: p63 might play an important role not only in squamous differentiation of lung cancer but also in tumor development and progression.

α粒子诱发BEP2D细胞转化过程中肺癌相关基因表达的cDNA Microarray研究

目的：探讨辐射诱发人支气管上皮细胞(BEP2D)转化过程中肺癌相关基因的表达。方法：用Cartesian PixSys5500 cDNA Microarray点样仪将60个肺癌相关基因以微阵列形式点布于醛基化的玻璃片上。提取α离子辐射前BEP2D细胞（原代）和α粒子辐射后20代、35代细胞总RNA，经长片段反转录和线性扩增标记成荧光探针后与微阵列中cDNA进行杂交。结果：原代细胞中检测到40个基因表达；20代检测到47个基因表达；35代检测到20个基因表达。所检测的基因中，抑癌基因的mRNA丰度在原代和20代后细胞中急剧下降；大多数癌基因的表达丰度在20代以后细胞中仅轻微下降；生长因子类基因大都在20代细胞表达。结论：在辐射诱发的人支气管上皮转化细胞中，抑癌基因的失活可能与细胞恶化有关；癌基因及生长因子类基因可能促进了细胞的转化。